CA1195931A - Method of stabilizing platelets for determining multiple platelet parameters in reference control and calibrator compositions; diluents therefor; and combination stabilization procedures - Google Patents
Method of stabilizing platelets for determining multiple platelet parameters in reference control and calibrator compositions; diluents therefor; and combination stabilization proceduresInfo
- Publication number
- CA1195931A CA1195931A CA000400904A CA400904A CA1195931A CA 1195931 A CA1195931 A CA 1195931A CA 000400904 A CA000400904 A CA 000400904A CA 400904 A CA400904 A CA 400904A CA 1195931 A CA1195931 A CA 1195931A
- Authority
- CA
- Canada
- Prior art keywords
- platelets
- acid
- platelet
- stabilizing
- iodoacetamide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/96—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood or serum control standard
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0226—Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2496/00—Reference solutions for assays of biological material
- G01N2496/05—Reference solutions for assays of biological material containing blood cells or plasma
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
- Y10T436/00—Chemistry: analytical and immunological testing
- Y10T436/10—Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
- Y10T436/101666—Particle count or volume standard or control [e.g., platelet count standards, etc.]
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
- Y10T436/00—Chemistry: analytical and immunological testing
- Y10T436/10—Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
- Y10T436/107497—Preparation composition [e.g., lysing or precipitation, etc.]
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
- Y10T436/00—Chemistry: analytical and immunological testing
- Y10T436/10—Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
- Y10T436/108331—Preservative, buffer, anticoagulant or diluent
Abstract
ABSTRACT
This invention utilizes for stabilization of platelets A
combination of iodoacetamide and an iminodiacetic acid or salt thereof, together with a compatible bacteriostatic agent, in an aqueous electrolytic solution which is maintained at a preselected range of pH and osmolality.
In a preferred formulation, the stabilizing solution contains iodoacetamide, N-(2-acetamido)iminodiacetic acid and sodium penicillin. Ethylenediaminetetraacetic acid, or salts thereof, are optionally present as an additional ingredient.
This invention utilizes for stabilization of platelets A
combination of iodoacetamide and an iminodiacetic acid or salt thereof, together with a compatible bacteriostatic agent, in an aqueous electrolytic solution which is maintained at a preselected range of pH and osmolality.
In a preferred formulation, the stabilizing solution contains iodoacetamide, N-(2-acetamido)iminodiacetic acid and sodium penicillin. Ethylenediaminetetraacetic acid, or salts thereof, are optionally present as an additional ingredient.
Description
s~
This invention concerns a method for ~tabilizing human or animal platelets ~lithout the use of aldehydes or other fixa~ive agents for determining multiple platelet parameters in reference coctrols using electronic instrumentation, as well as diluents therefor. As a result oE modern technological advances, instrumentation syscems for enumeration of cellular biological components for clinical diagnostic purposes has increased, and this has created a need for improvement of quality control productsO
To make the platelet referenc~ controls, the platelets are removed from the blood by centrifugation, washed with buffered salirle, and then "fixed" with agents such as glutaraldehyde, formaldehyde, pyruvic aldehyde, or the like. The aldehyde-reacted platelets are then su3pended in a buffer.
The s~lspended platelets are noe stable and will aggregate under a number of circumstarlces. In addition, the aldehyde-reacted platelets gradually change shape, shrinking in size as they age.
The aldehyde-reacted platelet3 are intended to be used in particle counters which are affected by particle ~i~e. Thu~, the reference control should ideally contain platelets which are as similar as possible in s;.~e to the platelet particles in normal human blooA. It is apparent that aggregation will affect the count and ~ize of the particles in the reference con~rols. It is also apparent that any shrinking or swelling which occurs also will impair the value of the control, since the platelets are no longer the sa~e size as pla~elets in fre~h human blood.
Se~eral human platelet reference controls are currently marketed. However, improveme~t is needed with respect to long term stability in the mea~urement of mean platelet volume and size distribution wid~h.
One such instru~eatatior~ sy~tem is the Coulter Counter~
Model S-Plus hematology system which counts and ~m;n~s the count and volume distribution of human platelets in whole blood specimen~.
Similar type instruments are manufactured by other companies for this purpose. Specifications as to electronic calibration are well defined to the operator of such in~trllments. Any human platelet reference control must satisfy all of the criteria that are measured on human patient specimens. ~he referecce con~rol needs to ~imulate as closely ~5~
as possible that of normal fresh whole blood specimens. It ~ust also conform to properly set thresholds as to the count mode, correctly balanced apertures, current, amplification gain setting~9 and responsiveness of the system as to all functional aspects.
Mean platelet volu~e distribution analysis has gained recognition recently as a useful measuremen~ for clinical application.
Human platele~ size distribution width can now be calculated from specifically measured parameters. Each of the measured parameters lends itself to improvement in eYamining the log normal distribution of human platelets. A1SOJ changeq and shif~s in the mode resul~ in erroneous counts.
The common cause of count discrepancies is aggregation.
Aggregatioll of plaeelets creates doublets that count in che white blood cell mode resulting in unreliable quality control measures. To the exten~ ~hat platelets undergo ag~regation, the aggreg~ted platelet cells appear and are counted aa white cells on the automated instrument. Since the number of platelets present are about 30 ~imes the number of white cells presen~ in fresh blood, even if only a small percentage of platelets aggregaee, this ean cause unreliable results with respect to the white cell count, especially after treatment with the usual fixing agents.
The adhesiveness of plate~ets also leads to false coltnts.
The adhesion of platelets to "foreign" surfaces has been measured by a variety of me~hods. ~he principle of all of these tes~s is the same.
The decrease i~ platelet count which occurs when blood is allowed to colltace a "foreign" surface for a standardized period of time is determined. Thi~t decrease ia indicative, in part, of the rlu~ber of platelets which have adhered to the foreign ~urface, a~d this value, expressed as a percentag~ of the original platelet count 9 has been called adhesiveness. However, it is known that some of the loss of platelets is also due to platele~ aggregaeion. Adhesiveness of platele~s, although dPpendent upon eitber calcium or magnesi~m ion s not as ~elective in this regard as the adhesiveness of th~
polymorphonucleocyt~s.
Platelets par~icipate in pri~ary hemostasis by forming aggregates at the site of the injured blood vessels. The agene ~hich is ultimately responsible for platelet aggrega~io~ is probably adenosine diph~sphate (ADP~, which may be derived from the injured 5~
tissues and erythrocytes or released from the platelets themselves by, among others, collagen, thrombin and epinephrine. In some patients with bleeding disorders, and in normal patients follow mg an ingestion of some drug~, platelet aggregation by one or more of these agents may be impaired. This impairment of platelet aggregation may be the cause of the prolonged bleeding time which i3 often obtained in these patients.
Ano~her discrepancy in platelet count is brought about by poor performance of stability until the expected expiration date of the control. Attempts to stabilize human platelets have proved ~o be extremely dificult. One major problem is that of disintegration of the platelec membrane. Fixatives have been used as a source of controlling disintegration. When platelet membranes disintegrate, they cause debris, thereby resulting in err~neous counts. New methods through improved computer technology for curve filling of raw data are beset by these problems.
In U.S. Patent 4,198,206 to Ryan (1980), a method is described for preparing a control from platelets which do not aggregate, and which have the same si~e as pla~ele~s in human blood, and maintain their si~e for at least 6 ~onths. ~liB control is a 3uspension of aldehyde-reacted platelets tha~ have been washed with a solution of:
(1) an a~ino acid which is glycine or alanine;
~2) glycol, glycerol or methanol;
(3) sodium chloride and sodium phospha~e; and (4) a solid polyethylene glycol (molecular weight 4000 to 20~000).
The propo~ed mechani3m of thi~ reaction show~ tha~ the a~ino group of the amino acid reacts with the aldehyde group so that further reaction, which includes cross-linking that leads ~o hardening and shrinking of the platelets, cannot occur.
The invention concerns a ~ethod of processing hu~an or animal platelets and stabili ing platele~s without the use of fixatives, such as glutaraldehyde, which lends itself to eliminate ~he afore~entioned discrepancies. Suprising and superior results are ob~ained by utili?il~g iodoa etamide, which has the for~ula 35~3~
ICH2CONH2, and ADA ~hich has the formula H2NCOCH2N(CH2COOH)2, without u8ing aldehyde-treated platelets. Long term cell stability is increased, without changing cell SiZ2 distribution.
This in~e~tion concerns a method for stabllizlng human or animal blood platelets without the use of aldehydes or o~her fixative agents for determining multiple platele~ parameters in reference controls uslng electronic instrumentation. More particularly, this inven~ion provides a novel stabilizing composition for suspending platelets which can act as a control reagent with ~he capaclty of monitoring the desired platelet parameters of a patlent's blood sample. These parameters include pletelet count, size distribution width, mean pla~elet volume3 and signal~noise ratio (debris), especlally in stand alone preparations.
Stabili~ation of platelets is then brought about by incorporating with an aqueous electrolyte solution in which the platelets are suspended a suitable quantity of a combination of ~1) iodoacetamide and (2) one or more substances selected from iminodiacetlc acids, and alkali metal and Alk~l~nP
buffer salts thereof, together with a compatible bacterios~atic agent, while maintaining the solution within a preselected range of pH and osmolality.
The preferred ~m~no~acetic acid is N-(2-acetamidojiminodiacetic acid (ADA) and the preferred bacteriostatic compound is a member of the pen~c~ 11 In family, especlally sodium penicillin, ~hich may also have other stabilizing action o~ the platelets.
One or more substances selected from ethylenp~minptetraacetic acid (EDTA), sodlum and potasslum salts thereof, and hydroxyethylethylene-diamine triacetic acid, may be included in the formulation.
This inventlon utili~es for stabillzation of platelets a c.o~binatlon of Lodoacetamide and an imlnoacetic acid or salt thereof, together wlth n compatible bacteriostatic agent, in an aqueous electrolytlc solution wlllch is maintained at a preselected range of pH and osmolality~
:` ``
3~
Iodoacetamide acts to reduce or inhibit adhesiveness of platelets and polymorphonuclear cells. Without being li~ited to any theory of action, it is known that platelets and polymorphonuclear cells have actively glycolyzing sys~ems, and data are consistent with the hypothesis that iodoacetamide acts on the adhesiveness by blocking glycolysis. However, data are like~ise consistent with any hypothesis which ascribes the adhes;veness to sul~hydryl dependent mechanisms with similar sensitivities to iodoacetamide.
The adhesiveness of platelets also appears to be dependent upon divalent cations, although the concentration required by the platelet~s seems to be much lo~er than that required by polymorphonuclear cells. Both magnesium and calcium ions are required for adhesiveness. Galcium ions alone are unable to restore adhesiveness to polymorphonuclear cells from blood which has been treated with a chelating resin to remove divalent cations.
The combined use oE N-(2-acetamido)iminodiacetic acid (ADA) and iodoacetamide has also shown that human platelets can be stabilized for extended periods of time without aggregation or losing their integrity. Iodoacetamide alone does not neutralize these factors. Also, buffering properties are not sufficient for suspendin~
in platelet rich plasma.
Combining iminodiacetic acid salts, especiaLly ADA, with iodoacetamide improves the stability, eliminates aggregation, and ~ines the hene~its of the anticoagulation properties of ADA. The 2S buf~ering propert;es of ADA are ideal for human platelets.
Approximately 0.5 g to about 2.0 g of iodoacetamide is added per liter of stabilizing solution.
Suitable iminodiacetic acid compounds are:
N-(2-acetamido)iminodiacetic acid (ADA)l sodium ADAJ potassium ADA, lithium ~DA, Tris-ADA or imidazole ADA. Approximately 0.5 g to about
This invention concerns a method for ~tabilizing human or animal platelets ~lithout the use of aldehydes or other fixa~ive agents for determining multiple platelet parameters in reference coctrols using electronic instrumentation, as well as diluents therefor. As a result oE modern technological advances, instrumentation syscems for enumeration of cellular biological components for clinical diagnostic purposes has increased, and this has created a need for improvement of quality control productsO
To make the platelet referenc~ controls, the platelets are removed from the blood by centrifugation, washed with buffered salirle, and then "fixed" with agents such as glutaraldehyde, formaldehyde, pyruvic aldehyde, or the like. The aldehyde-reacted platelets are then su3pended in a buffer.
The s~lspended platelets are noe stable and will aggregate under a number of circumstarlces. In addition, the aldehyde-reacted platelets gradually change shape, shrinking in size as they age.
The aldehyde-reacted platelet3 are intended to be used in particle counters which are affected by particle ~i~e. Thu~, the reference control should ideally contain platelets which are as similar as possible in s;.~e to the platelet particles in normal human blooA. It is apparent that aggregation will affect the count and ~ize of the particles in the reference con~rols. It is also apparent that any shrinking or swelling which occurs also will impair the value of the control, since the platelets are no longer the sa~e size as pla~elets in fre~h human blood.
Se~eral human platelet reference controls are currently marketed. However, improveme~t is needed with respect to long term stability in the mea~urement of mean platelet volume and size distribution wid~h.
One such instru~eatatior~ sy~tem is the Coulter Counter~
Model S-Plus hematology system which counts and ~m;n~s the count and volume distribution of human platelets in whole blood specimen~.
Similar type instruments are manufactured by other companies for this purpose. Specifications as to electronic calibration are well defined to the operator of such in~trllments. Any human platelet reference control must satisfy all of the criteria that are measured on human patient specimens. ~he referecce con~rol needs to ~imulate as closely ~5~
as possible that of normal fresh whole blood specimens. It ~ust also conform to properly set thresholds as to the count mode, correctly balanced apertures, current, amplification gain setting~9 and responsiveness of the system as to all functional aspects.
Mean platelet volu~e distribution analysis has gained recognition recently as a useful measuremen~ for clinical application.
Human platele~ size distribution width can now be calculated from specifically measured parameters. Each of the measured parameters lends itself to improvement in eYamining the log normal distribution of human platelets. A1SOJ changeq and shif~s in the mode resul~ in erroneous counts.
The common cause of count discrepancies is aggregation.
Aggregatioll of plaeelets creates doublets that count in che white blood cell mode resulting in unreliable quality control measures. To the exten~ ~hat platelets undergo ag~regation, the aggreg~ted platelet cells appear and are counted aa white cells on the automated instrument. Since the number of platelets present are about 30 ~imes the number of white cells presen~ in fresh blood, even if only a small percentage of platelets aggregaee, this ean cause unreliable results with respect to the white cell count, especially after treatment with the usual fixing agents.
The adhesiveness of plate~ets also leads to false coltnts.
The adhesion of platelets to "foreign" surfaces has been measured by a variety of me~hods. ~he principle of all of these tes~s is the same.
The decrease i~ platelet count which occurs when blood is allowed to colltace a "foreign" surface for a standardized period of time is determined. Thi~t decrease ia indicative, in part, of the rlu~ber of platelets which have adhered to the foreign ~urface, a~d this value, expressed as a percentag~ of the original platelet count 9 has been called adhesiveness. However, it is known that some of the loss of platelets is also due to platele~ aggregaeion. Adhesiveness of platele~s, although dPpendent upon eitber calcium or magnesi~m ion s not as ~elective in this regard as the adhesiveness of th~
polymorphonucleocyt~s.
Platelets par~icipate in pri~ary hemostasis by forming aggregates at the site of the injured blood vessels. The agene ~hich is ultimately responsible for platelet aggrega~io~ is probably adenosine diph~sphate (ADP~, which may be derived from the injured 5~
tissues and erythrocytes or released from the platelets themselves by, among others, collagen, thrombin and epinephrine. In some patients with bleeding disorders, and in normal patients follow mg an ingestion of some drug~, platelet aggregation by one or more of these agents may be impaired. This impairment of platelet aggregation may be the cause of the prolonged bleeding time which i3 often obtained in these patients.
Ano~her discrepancy in platelet count is brought about by poor performance of stability until the expected expiration date of the control. Attempts to stabilize human platelets have proved ~o be extremely dificult. One major problem is that of disintegration of the platelec membrane. Fixatives have been used as a source of controlling disintegration. When platelet membranes disintegrate, they cause debris, thereby resulting in err~neous counts. New methods through improved computer technology for curve filling of raw data are beset by these problems.
In U.S. Patent 4,198,206 to Ryan (1980), a method is described for preparing a control from platelets which do not aggregate, and which have the same si~e as pla~ele~s in human blood, and maintain their si~e for at least 6 ~onths. ~liB control is a 3uspension of aldehyde-reacted platelets tha~ have been washed with a solution of:
(1) an a~ino acid which is glycine or alanine;
~2) glycol, glycerol or methanol;
(3) sodium chloride and sodium phospha~e; and (4) a solid polyethylene glycol (molecular weight 4000 to 20~000).
The propo~ed mechani3m of thi~ reaction show~ tha~ the a~ino group of the amino acid reacts with the aldehyde group so that further reaction, which includes cross-linking that leads ~o hardening and shrinking of the platelets, cannot occur.
The invention concerns a ~ethod of processing hu~an or animal platelets and stabili ing platele~s without the use of fixatives, such as glutaraldehyde, which lends itself to eliminate ~he afore~entioned discrepancies. Suprising and superior results are ob~ained by utili?il~g iodoa etamide, which has the for~ula 35~3~
ICH2CONH2, and ADA ~hich has the formula H2NCOCH2N(CH2COOH)2, without u8ing aldehyde-treated platelets. Long term cell stability is increased, without changing cell SiZ2 distribution.
This in~e~tion concerns a method for stabllizlng human or animal blood platelets without the use of aldehydes or o~her fixative agents for determining multiple platele~ parameters in reference controls uslng electronic instrumentation. More particularly, this inven~ion provides a novel stabilizing composition for suspending platelets which can act as a control reagent with ~he capaclty of monitoring the desired platelet parameters of a patlent's blood sample. These parameters include pletelet count, size distribution width, mean pla~elet volume3 and signal~noise ratio (debris), especlally in stand alone preparations.
Stabili~ation of platelets is then brought about by incorporating with an aqueous electrolyte solution in which the platelets are suspended a suitable quantity of a combination of ~1) iodoacetamide and (2) one or more substances selected from iminodiacetlc acids, and alkali metal and Alk~l~nP
buffer salts thereof, together with a compatible bacterios~atic agent, while maintaining the solution within a preselected range of pH and osmolality.
The preferred ~m~no~acetic acid is N-(2-acetamidojiminodiacetic acid (ADA) and the preferred bacteriostatic compound is a member of the pen~c~ 11 In family, especlally sodium penicillin, ~hich may also have other stabilizing action o~ the platelets.
One or more substances selected from ethylenp~minptetraacetic acid (EDTA), sodlum and potasslum salts thereof, and hydroxyethylethylene-diamine triacetic acid, may be included in the formulation.
This inventlon utili~es for stabillzation of platelets a c.o~binatlon of Lodoacetamide and an imlnoacetic acid or salt thereof, together wlth n compatible bacteriostatic agent, in an aqueous electrolytlc solution wlllch is maintained at a preselected range of pH and osmolality~
:` ``
3~
Iodoacetamide acts to reduce or inhibit adhesiveness of platelets and polymorphonuclear cells. Without being li~ited to any theory of action, it is known that platelets and polymorphonuclear cells have actively glycolyzing sys~ems, and data are consistent with the hypothesis that iodoacetamide acts on the adhesiveness by blocking glycolysis. However, data are like~ise consistent with any hypothesis which ascribes the adhes;veness to sul~hydryl dependent mechanisms with similar sensitivities to iodoacetamide.
The adhesiveness of platelets also appears to be dependent upon divalent cations, although the concentration required by the platelet~s seems to be much lo~er than that required by polymorphonuclear cells. Both magnesium and calcium ions are required for adhesiveness. Galcium ions alone are unable to restore adhesiveness to polymorphonuclear cells from blood which has been treated with a chelating resin to remove divalent cations.
The combined use oE N-(2-acetamido)iminodiacetic acid (ADA) and iodoacetamide has also shown that human platelets can be stabilized for extended periods of time without aggregation or losing their integrity. Iodoacetamide alone does not neutralize these factors. Also, buffering properties are not sufficient for suspendin~
in platelet rich plasma.
Combining iminodiacetic acid salts, especiaLly ADA, with iodoacetamide improves the stability, eliminates aggregation, and ~ines the hene~its of the anticoagulation properties of ADA. The 2S buf~ering propert;es of ADA are ideal for human platelets.
Approximately 0.5 g to about 2.0 g of iodoacetamide is added per liter of stabilizing solution.
Suitable iminodiacetic acid compounds are:
N-(2-acetamido)iminodiacetic acid (ADA)l sodium ADAJ potassium ADA, lithium ~DA, Tris-ADA or imidazole ADA. Approximately 0.5 g to about
2.0 g o~ ADA, or an equivalent weight of a salt thereo-~, i6 added per liter of aqueous suspension oE the platelets, for example~ to platelet rich plasma.
The compatible bacteriostatic agent includes sodium penicillin G, potassium penioillin G, procaine penicillin G, penicillin V potassium, ampicillin and carbenicillin.
The compatible bacteriostatic agent includes sodium penicillin G, potassium penioillin G, procaine penicillin G, penicillin V potassium, ampicillin and carbenicillin.
3:~
PREFERRED EORMnLATIO~
1. Iodoacetamide 1.0 g 2. N-(2-acetamido)iminodiacetic acid (ADA) 1.0 g 3. Sodium chloride 9.416S g
PREFERRED EORMnLATIO~
1. Iodoacetamide 1.0 g 2. N-(2-acetamido)iminodiacetic acid (ADA) 1.0 g 3. Sodium chloride 9.416S g
4. Sodium penicillin 0.155 g q.s. ~o l/Liter with distilled water Osmolality = 310 Adjust pH to 7.0 ~ 0.1 with NaOH.
In the above for~ulation, it is desirable to add ethylenediaminetetraacetic acid (~DTA), or salts thereof, as an optionsl ingredient to act as a chelating agent for divalent ions.
Approximately 0.5 g to about 2.0 g of EDTA, or an equi~alent weigh~ of ialt thereof, is added per liter of aqueou~ suspension of the platelets.
Method of Preparation 1. Add approxima~ely 500 ~1 distilled water to a 1 liter flask.
2. Add ADA 1 g and allow to dissolve.
3. When disodium EDTA is included in ~he formulation, add 1 g and allo~ to dissolve.
4. Add iodoacetamide 1 g and allow to dissolve.
In the above for~ulation, it is desirable to add ethylenediaminetetraacetic acid (~DTA), or salts thereof, as an optionsl ingredient to act as a chelating agent for divalent ions.
Approximately 0.5 g to about 2.0 g of EDTA, or an equi~alent weigh~ of ialt thereof, is added per liter of aqueou~ suspension of the platelets.
Method of Preparation 1. Add approxima~ely 500 ~1 distilled water to a 1 liter flask.
2. Add ADA 1 g and allow to dissolve.
3. When disodium EDTA is included in ~he formulation, add 1 g and allo~ to dissolve.
4. Add iodoacetamide 1 g and allow to dissolve.
5. Add sodium chloride 9.4166 g and allow to dissolve.
6. Add ~odium penicillin 0.155 g and allow to dissolve.
7. q. 9 . ~ith distilled water to 1 liter.
3. Adju~t pH to 7.0 ~ 0.5 with sodium hydroxide, and osmolality of ~5 33~ ~ 30 with sodiu~ chloride.
9. Filter through 0.22 micron filter into sterile bags.
10. Store at room temperaeure up to 9iX mon~hs.
Jaeger U.S. Patent 4,116,635 s~udied the an~icoagulznt properties of both EDTA and ~DA using whole blood, and reported the results in TABLE I cf his patent spscification. Log K', which is the conditional ~tability coll3tant at p~ of 7.5, wa~ Eound to be 7.9 or EDTA and only 4.0 for ADA. In a proce3~ for testing a sample of blood for coagulation and ~ther factors according eo his Claim 19 the anticoagulent ielected had a Log K' from 3.4 to about 4.2, ~hus including ADA, but excluding EDTAo 3 ~
IL ~ .
Lt is suprising that, in our invention, best results are obtained when both ADA and EDTA are used together in approximately equal amounts in the for~ulation. The amount of EDTA which is added to the aqueous formulation of platelets is approximately 0.5 g to about 2.0 g per liter.
The following examples illustrate the ~ethod of stabilizing blood pla~elets without the use of fixative agents for determining multiple platelet para~eters by electronic inst~umentation.
STABILITY OF PLATELET RICH PLASMA WITHOUT
TR~ATM~.NT WITH STABILIZING SOLUTION
Time S/N MPV
initial 5.55 6.6u after 4 days 5.00 6.6u after 4 days 2.08 6.1u after 6 days 1.82 6.0u after 7 days 1.43 no it The above resul~s were ob~ained when stabilizing solution was stored at 2 to 8~C and assayed on a standard Coulter Counter~
` ~, Model S-Plus instrument. ~/~
The signal/noise ra~io~decreased with time, i.e., the noise increased relative to the signal. All parameters exhibited changes with ti~e. After seven day~ ! complete cellular fragmentation wa~
demonstra~ed, resulting in a no-fi~ mode of the distributioLI analysis char~.. The platelet count, mean platelet volume ~MPV~, and si~e distribution could not be determined by the instrument.
Representative samples of gluearaldehyde fixed platelets in phosphate buffered sali~e solution likewi~e sho~ a decrease in mean platelet volume over a period of ti~e. Currently manufactured clinical hema~ological instrumentation cannot tGlerate a shift ~o the left (decrease) of the mean platelet si~,e distribution. This shift re~ults in a no~fit condition.
Excessive debris produces no-fit in size dis~ribution curve~.
~9~
REPR~SENTATIVE SAMPLES AFTER TRFAT~NT OF PLATELETS WITH
STABILIZING SOLUTIQN HAVING THE PREFERRED FORMNLATION
Mean Mean Day No. S/~ Coun~Platele~ Vol. Size Dis~ribution 4.54 206 7.~ 17.1 4.0 202 7.9 17.1 4.0 193 7.6 17.1 120 4.0 206 7.7 17.2 180 4.0 193 7.6 17.1 The abQve results were ob~ained when stored at 2 to 8C and assayed on a standard Coulter Counter~ Model S-Plus instrument.
These re~ults sho~ that there is no significant chan~e in any of the parameters after 180 days. There is excellent stability ~ith re~pect to all critical par~meters necessary for quality control applications measured by electronic devices.
Signal/noise ratios mu9t be maximized to assure the operator that the instrument i8 perfor~ing according to the manufacturer'2 specifications. Increased noise has been an ever presene problem because of platelet debri~ and cellular fragmentation during the life o such products. In these sa~ples 9 the signal/noise ratio shows no decrease, which would render the in~rument incapable of critically measuring parameeers for quality control measures. Decreased ~;gnal/noise ratios on electronic particle sizing de~ices, e~pecially in the lo~er thresholds where platelets are coun~ed and their size distribution deterTnined, cannot be tolerated.
Statistical data document~ the use oE the above stabilized human platelets in a whole blood control preparation beyond 100 days in several clinical hematology laboratorie~.
Preconditioning hu~an platelet~ with the stabili~ing solution of this invention allows the cells to be resuspended into a hu~an or animal red blo~d cell preparatlon as a con~rol For ~oni~oring electronic devices. Al~o, it i8 pO8 ible to stabiliæe hu~an piatelets for in vitro therapeutic applications.
For praconditioning the pl~telets~ approxima~ely 50 to 100 ml of the stabilizing diluent ia added directly to a 200 ~ 50 g uni~
of platelet rich plasma (PRP) and mixed well. The stabilized PRP can then stand at room temperature Eor 7 days before cen~rifugation for 3:~
procurement of platelet concentrate. In the alternative! the PRP with addition of stabilizing diluent may be stored at 4 to 6C for up to 3 months before processing by centrifu~ation to harves~ the platelet concentrateO
PRP i8 centrifuced at approximately 3,800 RPM for 5 minutes (refrigerated centrifuge~ and the supernate is expressed off to waste.
Phosphate buffered saline at pH 7.0 to 7.2 and 320 mOs/kg is added with mixing and the washing step is repeated a second time. The platelets are concentra~ed after the seco~d wash and pooled into a single high concentration pool.
The platelet concentra~e is resuspended in the diluent at ~he desired concentration, and this preparation is added to stabilized erythrocytes to be utilized as a whole blood control. The final preparation i~ stored at 2 to 8C for up to 6 months.
This diluent successfully stabilizes platelet rich plasma procured with the use of commonly utilized anticoagulents such as CPD
(citrate-phosphate-dextrose), ACD (acid-ci~rate-dextrose) and EDTA
(ethylenediaminetetraacetic acid).
The s~abilized platelets can be utilized in stand alone platelet controls, a3 well as in whole blood reference controls, to as~ure good quality control procedures and monitor multiple platelet parameters including (1) count, (2) mean size distribution, (3) mean platelet volume, and (4) signal/noise ratio (debriR).
It is well recognized in the art that stabilized blood cell preparations useful for checking the accuracy of au~omatic counting instruments for blood components are subject to some degree of acceptable variation. Such preparations are discussed briefly on page 14 of the book HEMATOLO~Y, by Williams9 Beutler, Er~lev, and Rundle~, McGraw~ Inc. (1972).
In the art of hematology~ a difference i8 made between the terms "calibrator" and ':reference control". Calibraeors are blood cell preparationR that are used to set the aueo~ated instrument with respect to certain key nu~bers, and are used only once. Reference controls are blood cell preparations used from time to time to check ehe continuing ac~uracy oE an aueo~ated instru~en~ which has previously been "calibrated" with the calibra~or blood cell preparation.
3~
, , It i~ an ad~antage of this in~ention that platelets which have been stabilized by the methods herein described, without the use of aldehydes as fixing agents to give reference controls which are stable for 60 days and longer, can then be subjected to a second stabilization treatment by other methods, including method~ employing aldehydes aDd/or surfactants to give platelet preparations having improved stability over a longer period of time, especially for certa;n purposes. Such preparations are particularly usef~l în calibrator compo~i~ion~.
Fo~ example, the platele~ stabilized by the preferred formulation of this invention can then be treated with a fixative~-stabilizer ~edia having the following formulation:
NaH2P04-H2o 0.196 g Na2HP04-7H20 0.98Q g NaN3 0.098 g NaCl 7.9 g G'lutaraldehyde, 49% 8.4 g Tergitol~ 15-S-12 0.5 g Water q.s. 1 L
Ad.just to p~ 7.3 to 7.4 wi~h phosphate, and to an osmolality of 290 mOs/kg with NaCl.
Tergitol~ 15-S-12 i~ a mixture of ethoxylates oE isomeric linear alcohols, as de~cri~ed in U.S. Patent 3,91~,450 (1975) and U.S.
Patent 3,968,248 (197~), Boucher to Wave Energy Sy~tems, Inc., and ~old as Wavecide~Ol. 'l'he mixture has the for~ula.
CH3-(GH?)n~C~3 O- ( CH~CH2-0 ) X-EI
in which the polyoxyethylene chain i~ randomly attached to the linear aliphatic chain, wherein n = 9 to 13 and x = 9 ~o 13.
3. Adju~t pH to 7.0 ~ 0.5 with sodium hydroxide, and osmolality of ~5 33~ ~ 30 with sodiu~ chloride.
9. Filter through 0.22 micron filter into sterile bags.
10. Store at room temperaeure up to 9iX mon~hs.
Jaeger U.S. Patent 4,116,635 s~udied the an~icoagulznt properties of both EDTA and ~DA using whole blood, and reported the results in TABLE I cf his patent spscification. Log K', which is the conditional ~tability coll3tant at p~ of 7.5, wa~ Eound to be 7.9 or EDTA and only 4.0 for ADA. In a proce3~ for testing a sample of blood for coagulation and ~ther factors according eo his Claim 19 the anticoagulent ielected had a Log K' from 3.4 to about 4.2, ~hus including ADA, but excluding EDTAo 3 ~
IL ~ .
Lt is suprising that, in our invention, best results are obtained when both ADA and EDTA are used together in approximately equal amounts in the for~ulation. The amount of EDTA which is added to the aqueous formulation of platelets is approximately 0.5 g to about 2.0 g per liter.
The following examples illustrate the ~ethod of stabilizing blood pla~elets without the use of fixative agents for determining multiple platelet para~eters by electronic inst~umentation.
STABILITY OF PLATELET RICH PLASMA WITHOUT
TR~ATM~.NT WITH STABILIZING SOLUTION
Time S/N MPV
initial 5.55 6.6u after 4 days 5.00 6.6u after 4 days 2.08 6.1u after 6 days 1.82 6.0u after 7 days 1.43 no it The above resul~s were ob~ained when stabilizing solution was stored at 2 to 8~C and assayed on a standard Coulter Counter~
` ~, Model S-Plus instrument. ~/~
The signal/noise ra~io~decreased with time, i.e., the noise increased relative to the signal. All parameters exhibited changes with ti~e. After seven day~ ! complete cellular fragmentation wa~
demonstra~ed, resulting in a no-fi~ mode of the distributioLI analysis char~.. The platelet count, mean platelet volume ~MPV~, and si~e distribution could not be determined by the instrument.
Representative samples of gluearaldehyde fixed platelets in phosphate buffered sali~e solution likewi~e sho~ a decrease in mean platelet volume over a period of ti~e. Currently manufactured clinical hema~ological instrumentation cannot tGlerate a shift ~o the left (decrease) of the mean platelet si~,e distribution. This shift re~ults in a no~fit condition.
Excessive debris produces no-fit in size dis~ribution curve~.
~9~
REPR~SENTATIVE SAMPLES AFTER TRFAT~NT OF PLATELETS WITH
STABILIZING SOLUTIQN HAVING THE PREFERRED FORMNLATION
Mean Mean Day No. S/~ Coun~Platele~ Vol. Size Dis~ribution 4.54 206 7.~ 17.1 4.0 202 7.9 17.1 4.0 193 7.6 17.1 120 4.0 206 7.7 17.2 180 4.0 193 7.6 17.1 The abQve results were ob~ained when stored at 2 to 8C and assayed on a standard Coulter Counter~ Model S-Plus instrument.
These re~ults sho~ that there is no significant chan~e in any of the parameters after 180 days. There is excellent stability ~ith re~pect to all critical par~meters necessary for quality control applications measured by electronic devices.
Signal/noise ratios mu9t be maximized to assure the operator that the instrument i8 perfor~ing according to the manufacturer'2 specifications. Increased noise has been an ever presene problem because of platelet debri~ and cellular fragmentation during the life o such products. In these sa~ples 9 the signal/noise ratio shows no decrease, which would render the in~rument incapable of critically measuring parameeers for quality control measures. Decreased ~;gnal/noise ratios on electronic particle sizing de~ices, e~pecially in the lo~er thresholds where platelets are coun~ed and their size distribution deterTnined, cannot be tolerated.
Statistical data document~ the use oE the above stabilized human platelets in a whole blood control preparation beyond 100 days in several clinical hematology laboratorie~.
Preconditioning hu~an platelet~ with the stabili~ing solution of this invention allows the cells to be resuspended into a hu~an or animal red blo~d cell preparatlon as a con~rol For ~oni~oring electronic devices. Al~o, it i8 pO8 ible to stabiliæe hu~an piatelets for in vitro therapeutic applications.
For praconditioning the pl~telets~ approxima~ely 50 to 100 ml of the stabilizing diluent ia added directly to a 200 ~ 50 g uni~
of platelet rich plasma (PRP) and mixed well. The stabilized PRP can then stand at room temperature Eor 7 days before cen~rifugation for 3:~
procurement of platelet concentrate. In the alternative! the PRP with addition of stabilizing diluent may be stored at 4 to 6C for up to 3 months before processing by centrifu~ation to harves~ the platelet concentrateO
PRP i8 centrifuced at approximately 3,800 RPM for 5 minutes (refrigerated centrifuge~ and the supernate is expressed off to waste.
Phosphate buffered saline at pH 7.0 to 7.2 and 320 mOs/kg is added with mixing and the washing step is repeated a second time. The platelets are concentra~ed after the seco~d wash and pooled into a single high concentration pool.
The platelet concentra~e is resuspended in the diluent at ~he desired concentration, and this preparation is added to stabilized erythrocytes to be utilized as a whole blood control. The final preparation i~ stored at 2 to 8C for up to 6 months.
This diluent successfully stabilizes platelet rich plasma procured with the use of commonly utilized anticoagulents such as CPD
(citrate-phosphate-dextrose), ACD (acid-ci~rate-dextrose) and EDTA
(ethylenediaminetetraacetic acid).
The s~abilized platelets can be utilized in stand alone platelet controls, a3 well as in whole blood reference controls, to as~ure good quality control procedures and monitor multiple platelet parameters including (1) count, (2) mean size distribution, (3) mean platelet volume, and (4) signal/noise ratio (debriR).
It is well recognized in the art that stabilized blood cell preparations useful for checking the accuracy of au~omatic counting instruments for blood components are subject to some degree of acceptable variation. Such preparations are discussed briefly on page 14 of the book HEMATOLO~Y, by Williams9 Beutler, Er~lev, and Rundle~, McGraw~ Inc. (1972).
In the art of hematology~ a difference i8 made between the terms "calibrator" and ':reference control". Calibraeors are blood cell preparationR that are used to set the aueo~ated instrument with respect to certain key nu~bers, and are used only once. Reference controls are blood cell preparations used from time to time to check ehe continuing ac~uracy oE an aueo~ated instru~en~ which has previously been "calibrated" with the calibra~or blood cell preparation.
3~
, , It i~ an ad~antage of this in~ention that platelets which have been stabilized by the methods herein described, without the use of aldehydes as fixing agents to give reference controls which are stable for 60 days and longer, can then be subjected to a second stabilization treatment by other methods, including method~ employing aldehydes aDd/or surfactants to give platelet preparations having improved stability over a longer period of time, especially for certa;n purposes. Such preparations are particularly usef~l în calibrator compo~i~ion~.
Fo~ example, the platele~ stabilized by the preferred formulation of this invention can then be treated with a fixative~-stabilizer ~edia having the following formulation:
NaH2P04-H2o 0.196 g Na2HP04-7H20 0.98Q g NaN3 0.098 g NaCl 7.9 g G'lutaraldehyde, 49% 8.4 g Tergitol~ 15-S-12 0.5 g Water q.s. 1 L
Ad.just to p~ 7.3 to 7.4 wi~h phosphate, and to an osmolality of 290 mOs/kg with NaCl.
Tergitol~ 15-S-12 i~ a mixture of ethoxylates oE isomeric linear alcohols, as de~cri~ed in U.S. Patent 3,91~,450 (1975) and U.S.
Patent 3,968,248 (197~), Boucher to Wave Energy Sy~tems, Inc., and ~old as Wavecide~Ol. 'l'he mixture has the for~ula.
CH3-(GH?)n~C~3 O- ( CH~CH2-0 ) X-EI
in which the polyoxyethylene chain i~ randomly attached to the linear aliphatic chain, wherein n = 9 to 13 and x = 9 ~o 13.
Claims (12)
PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:
1. A method of stabilizing platelets comprising:
incorporating with platelets in an aqueous electrolyte suspending fluid a stabilizing solution which is an aqueous solution of 1): iodoacetamide and 2): one or more substances selected from iminodiacetic acids and alkali metal salts and alkaline buffer salts of said acid, together with a compatible bacteriostatic agent, wherein the resultant media is maintained at a preselected range of pH and osmolality, for example, using an alkali metal hydroxide and alkali metal chloride.
incorporating with platelets in an aqueous electrolyte suspending fluid a stabilizing solution which is an aqueous solution of 1): iodoacetamide and 2): one or more substances selected from iminodiacetic acids and alkali metal salts and alkaline buffer salts of said acid, together with a compatible bacteriostatic agent, wherein the resultant media is maintained at a preselected range of pH and osmolality, for example, using an alkali metal hydroxide and alkali metal chloride.
2. The method of claim 1 further comprising: causing the platelets to stand for a suitable time and temperature to achieve at least partial stabilization, separating the platelets from said stabilizing solution; and adding thereto a suitable quantity of glutaraldehyde and a non-ionic surfactant which is a mixture of ethoxylates of isomeric linear alcohols having the formula:
wherein the polyoxyethylene chain is randomly attached to the linear aliphatic chain, and wherein n = 9 to 13 and x = 9 to 13, said composition being adjusted to a preselected range of pH and osmolality.
wherein the polyoxyethylene chain is randomly attached to the linear aliphatic chain, and wherein n = 9 to 13 and x = 9 to 13, said composition being adjusted to a preselected range of pH and osmolality.
3. The method of claim 1 wherein ethylenediaminetetraacetic acid, the sodium or potassium salts thereof, is present as an additional ingredient of the stabilizing solution.
4. The method of any one of claims 1, 2 or 3 wherein said iminodiacetic acid is N-(2-acetamido)iminodiacetic acid.
5. The method of any one of claims 1, 2 or 3 wherein said iminodiacetic acid is present in a concentration of 0.5 g to 2.0 g per liter.
6. The method of any one of claims 1, 2 or 3 wherein said bacteriostatic agent is a member of the penicillin family comprising: penicillin G, potassium penicillin G, procaine penicillin G, penicillin V potassium, ampicillin and carbenicillin.
7. The method of any one of claims 1, 2 or 3 wherein said stabilizing solution is added to platelet rich plasma.
8. The method of any one of claim 1, 2 or 3 wherein said iodoacetamide is present in a concentration of 0.5 g to 2.0 g per liter.
9. A diluent suitable for use in electronic instrumentation for stabilizing platelets for determining multiple platelet parameters, said diluent comprising: an aqueous solution of iodoacetamide and an iminodiacetic acid, and the alkali metal and alkaline buffer salts of said acid, together with compatible bacteriostatic agents; said diluent being an aqueous electrolytic solution maintained within a preselected range of pH and osmolality, for furnishing long term platelet stability and freedom from particulate matter which causes erroneous results.
10. A hemotology reference control media for determining multiple platelet parameters utilizing electronic instrumentation, said media comprising a suspension of platelets in an aqueous solution of iodoacetamide and iminodiacetic said, or the alkali metal or alkaline buffer salts of said acid, together with a compatible bacteriostatic agent, wherein the resultant media is maintained at a preselected range of pH and osmolality.
11. The media of claim 10 wherein ethylenediaminetetraacetic acid, or the sodium or potassium salts thereof, is an additional ingredient of the stabilizing solution.
12. A multiple analysis hematology reference control media comprising the control media of claim 10 or 11 which is capable of being used for testing the accuracy and precision of classical red blood cell and platelet parameters.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US06/268,050 US4405719A (en) | 1981-05-29 | 1981-05-29 | Method of stabilizing platelets for determining multiple platelet parameters in reference control and calibrator compositions; diluents therefor; and combination stabilization procedures |
| US06/268,050 | 1981-05-29 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1195931A true CA1195931A (en) | 1985-10-29 |
Family
ID=23021258
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000400904A Expired CA1195931A (en) | 1981-05-29 | 1982-04-13 | Method of stabilizing platelets for determining multiple platelet parameters in reference control and calibrator compositions; diluents therefor; and combination stabilization procedures |
Country Status (8)
| Country | Link |
|---|---|
| US (1) | US4405719A (en) |
| JP (1) | JPS57203957A (en) |
| CA (1) | CA1195931A (en) |
| DE (1) | DE3220232A1 (en) |
| ES (1) | ES8400874A1 (en) |
| FR (1) | FR2506945A1 (en) |
| GB (1) | GB2099281B (en) |
| HK (1) | HK10686A (en) |
Families Citing this family (26)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA1216518A (en) * | 1982-11-01 | 1987-01-13 | Gail A. Rock | Plasma-free medium for platelet storage |
| US4471051A (en) * | 1983-06-13 | 1984-09-11 | New England Medical Center Hospitals, Inc. | Stabilization of neutrophils and platelets |
| US4828976A (en) * | 1983-12-29 | 1989-05-09 | Thomas Jefferson University | Glucose free media for storing blood platelets |
| US4704364A (en) * | 1984-05-18 | 1987-11-03 | Coulter Electronics, Inc. | Hematology control compositions for three populations of leukocytes; and methods for their preparation and use in whole blood control systems |
| US4847204A (en) * | 1984-05-24 | 1989-07-11 | Southeast Vetlab, Inc. | Calibrator composition and method of producing and using same for veterinary applications |
| US4711852A (en) * | 1984-11-05 | 1987-12-08 | Akzo N.V. | Control for blood gas analyzers and hemoglobin analysis |
| US4858154A (en) * | 1986-04-07 | 1989-08-15 | Coulter Electronics, Inc. | Interlaboratory quality assurance program |
| US4788139A (en) * | 1987-08-06 | 1988-11-29 | Streck Laboratories, Inc. | Platelet aggregation reagent, reagent container and method of determining platelet aggregation in EDTA-anticoagulated blood |
| US5008202A (en) * | 1988-11-29 | 1991-04-16 | Sequoia Turner Corporation | Blood diluent for red blood cell analysis |
| DE3938907C2 (en) * | 1989-11-24 | 1999-11-04 | Dade Behring Marburg Gmbh | Means for storing and suspending cells, in particular erythrocytes |
| US5256571A (en) * | 1991-05-01 | 1993-10-26 | Cytyc Corporation | Cell preservative solution |
| US6509192B1 (en) | 1992-02-24 | 2003-01-21 | Coulter International Corp. | Quality control method |
| US6362003B1 (en) | 1992-02-24 | 2002-03-26 | Coulter Corporation | Hematological reference control composition containing leukocyte analogs, methods of making, and uses thereof |
| EP0721104A2 (en) * | 1993-07-05 | 1996-07-10 | Northern General Hospital N.H.S. Trust | Preparation and stabilisation of cells |
| US6197540B1 (en) * | 1993-07-05 | 2001-03-06 | Northern General Hospital N.H.S. Trust | Preparation and stabilization of cells using aged transition metal solutions |
| CN1149338A (en) * | 1994-04-05 | 1997-05-07 | 北方总医院N·H·S·托拉斯 | Preparation and stabilisation of cell suspensions |
| JPH09509681A (en) | 1994-12-16 | 1997-09-30 | バクスター、インターナショナル、インコーポレイテッド | Control of donor blood characteristics |
| DE19634313A1 (en) * | 1996-08-24 | 1998-02-26 | Behringwerke Ag | Method for stabilizing platelets |
| US7198953B2 (en) * | 2003-10-12 | 2007-04-03 | Beckman Coulter, Inc. | Method of using a reference control composition for measurement of nucleated red blood cells |
| US7135341B2 (en) * | 2004-04-07 | 2006-11-14 | Beckman Coulter, Inc. | Reference control containing a nucleated red blood cell component |
| US7109036B2 (en) * | 2004-05-13 | 2006-09-19 | Beckman Coulter, Inc. | Hematology reference control containing an immature granulocyte component |
| US7531357B2 (en) * | 2005-04-04 | 2009-05-12 | Bio-Rad Laboratories, Inc. | Preparation of platelet analogs |
| US7354767B2 (en) * | 2006-03-16 | 2008-04-08 | Beckman Coulter, Inc. | Reference control composition containing a nucleated red blood cell component made of non-nucleated blood cells |
| US7754487B2 (en) * | 2006-11-14 | 2010-07-13 | Beckman Coulter, Inc. | Hematology linearity control composition, system and method of use |
| US8835104B2 (en) | 2007-12-20 | 2014-09-16 | Fenwal, Inc. | Medium and methods for the storage of platelets |
| EP2694131B1 (en) | 2011-04-07 | 2019-08-28 | Fenwal, Inc. | Automated methods and systems for providing platelet concentrates with reduced residual plasma volumes and storage media for such platelet concentrates |
Family Cites Families (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3968250A (en) * | 1971-06-21 | 1976-07-06 | Wave Energy Systems, Inc. | Method and sporicidal compositions for synergistic disinfection or sterilization |
| US3873467A (en) * | 1974-02-01 | 1975-03-25 | United Medical Lab Inc | Hematologic reference control |
| US3962125A (en) * | 1975-01-13 | 1976-06-08 | Coulter Diagnostics, Inc. | Multi-purpose diluent for use in blood analysis by electronic instrumentation of the coulter type |
| DE2546166A1 (en) * | 1975-10-15 | 1977-04-28 | Behringwerke Ag | TANNED THROMBOCYTE |
| US4116635A (en) * | 1977-03-14 | 1978-09-26 | Jaeger Mark A | General purpose in vitro anticoagulant |
| US4160644A (en) * | 1977-06-13 | 1979-07-10 | Streck Laboratories, Inc. | Platelet reference control and method of preparation |
| US4198206A (en) * | 1977-06-13 | 1980-04-15 | Ryan Wayne L | Method for preparing a platelet reference control |
| US4302355A (en) * | 1977-08-01 | 1981-11-24 | Warner-Lambert Company | Platelet reference control |
| US4213876A (en) * | 1978-08-22 | 1980-07-22 | Coulter Electronics, Inc. | Multi-purpose blood diluent for use in electronic blood analysis instrumentation |
| US4299726A (en) * | 1979-05-07 | 1981-11-10 | Coulter Electronics, Inc. | Process for preparing whole blood reference controls having long term stability, preconditioning diluent and media therefor |
| US4324686A (en) * | 1979-06-11 | 1982-04-13 | R & D Systems, Inc. | Hematology control composition and methods for its use |
-
1981
- 1981-05-29 US US06/268,050 patent/US4405719A/en not_active Expired - Fee Related
-
1982
- 1982-04-13 CA CA000400904A patent/CA1195931A/en not_active Expired
- 1982-05-28 ES ES512661A patent/ES8400874A1/en not_active Expired
- 1982-05-28 DE DE19823220232 patent/DE3220232A1/en active Granted
- 1982-05-28 GB GB8215792A patent/GB2099281B/en not_active Expired
- 1982-05-28 JP JP57090032A patent/JPS57203957A/en active Granted
- 1982-05-28 FR FR8209414A patent/FR2506945A1/en active Granted
-
1986
- 1986-02-13 HK HK106/86A patent/HK10686A/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| ES512661A0 (en) | 1983-12-01 |
| GB2099281A (en) | 1982-12-08 |
| ES8400874A1 (en) | 1983-12-01 |
| JPS57203957A (en) | 1982-12-14 |
| JPS648306B2 (en) | 1989-02-13 |
| DE3220232C2 (en) | 1987-10-22 |
| DE3220232A1 (en) | 1982-12-23 |
| FR2506945A1 (en) | 1982-12-03 |
| US4405719A (en) | 1983-09-20 |
| GB2099281B (en) | 1985-05-22 |
| HK10686A (en) | 1986-02-21 |
| FR2506945B3 (en) | 1984-04-13 |
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