WO2007015906A3 - Methods and compositions for smooth muscle reconstruction - Google Patents
Methods and compositions for smooth muscle reconstruction Download PDFInfo
- Publication number
- WO2007015906A3 WO2007015906A3 PCT/US2006/028040 US2006028040W WO2007015906A3 WO 2007015906 A3 WO2007015906 A3 WO 2007015906A3 US 2006028040 W US2006028040 W US 2006028040W WO 2007015906 A3 WO2007015906 A3 WO 2007015906A3
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- WO
- WIPO (PCT)
- Prior art keywords
- smooth muscle
- population
- composition
- compositions
- provides
- Prior art date
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-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
- C12N5/0667—Adipose-derived stem cells [ADSC]; Adipose stromal stem cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
Abstract
This invention also provides a purified or isolated population of ADSCs that can differentiate into a cell of the leiomyogenic lineage, e.g., smooth muscle or skeletal muscle. In yet another aspect, the population additionally can be differentiated into a lineage selected from the group consisting of osteogenic, adipogenic, chondrogenic, myogenic and neuronal. This invention further provides a composition comprising a substantially homogeneous expanded population of smooth muscle cells. This invention provides a composition comprising a substantially homogeneous expanded population of skeletal muscle cells. Also provided herein is an isolated composition comprising a purified adipose-derived stem cell (ADSC) or progeny of said ADSC and an effective amount of laminin or heparin, effective to induce leiomyogenic differentiation. Diagnostic and therapeutic uses for these compositions are provided herein.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US11/192,753 | 2005-07-29 | ||
US11/192,753 US7531355B2 (en) | 2005-07-29 | 2005-07-29 | Methods and compositions for smooth muscle reconstruction |
Publications (2)
Publication Number | Publication Date |
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WO2007015906A2 WO2007015906A2 (en) | 2007-02-08 |
WO2007015906A3 true WO2007015906A3 (en) | 2009-04-16 |
Family
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Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US2006/028040 WO2007015906A2 (en) | 2005-07-29 | 2006-07-18 | Methods and compositions for smooth muscle reconstruction |
Country Status (2)
Country | Link |
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US (1) | US7531355B2 (en) |
WO (1) | WO2007015906A2 (en) |
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US8313954B2 (en) | 2009-04-03 | 2012-11-20 | Biomet Biologics, Llc | All-in-one means of separating blood components |
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US9352003B1 (en) | 2010-05-14 | 2016-05-31 | Musculoskeletal Transplant Foundation | Tissue-derived tissuegenic implants, and methods of fabricating and using same |
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US9296984B2 (en) | 2010-07-09 | 2016-03-29 | The Gid Group, Inc. | Tissue processing apparatus and method for processing adipose tissue |
WO2014039697A1 (en) | 2012-09-06 | 2014-03-13 | The Gid Group, Inc. | Tissue processing apparatus and method for processing adipose tissue |
US9206387B2 (en) | 2010-07-09 | 2015-12-08 | The Gid Group, Inc. | Method and apparatus for processing adipose tissue |
EP3366327B1 (en) | 2010-07-09 | 2022-09-21 | GID BIO, Inc. | Apparatus and methods relating to collecting and processing human biological material containing adipose |
WO2013106655A1 (en) | 2012-01-11 | 2013-07-18 | The Gid Group, Inc. | Method for processing adipose tissue and processing apparatus |
CN103443272A (en) | 2010-12-27 | 2013-12-11 | 智能细胞生物科学有限公司 | Ultrasonic cavitation derived stromal or mesenchymal vascular extracts and cells derived therefrom obtained from adipose tissue and use thereof |
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4443546A (en) * | 1980-07-07 | 1984-04-17 | The Beth Israel Hospital Association | Process and composition for propagating mammalian cells |
US5605687A (en) * | 1992-05-15 | 1997-02-25 | Arch Development Corporation | Methods and compositions of a polymer (poloxamer) for repair of electrical injury |
US20030161816A1 (en) * | 2001-12-07 | 2003-08-28 | Fraser John K. | Systems and methods for treating patients with processed lipoaspirate cells |
US20040193274A1 (en) * | 2003-03-28 | 2004-09-30 | Trieu Hai H. | Materials and methods for augmenting and/or repairing intervertebral discs |
Family Cites Families (32)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU713280B2 (en) | 1995-11-16 | 1999-11-25 | Case Western Reserve University | In vitro chondrogenic induction of human mesenchymal stem cells |
US6200606B1 (en) | 1996-01-16 | 2001-03-13 | Depuy Orthopaedics, Inc. | Isolation of precursor cells from hematopoietic and nonhematopoietic tissues and their use in vivo bone and cartilage regeneration |
AU2730497A (en) | 1996-04-17 | 1997-11-07 | Case Western Reserve University | Cryopreservation and extensive subculturing of human mesenchymal stem cells |
WO1997040137A1 (en) | 1996-04-19 | 1997-10-30 | Osiris Therapeutics, Inc. | Regeneration and augmentation of bone using mesenchymal stem cells |
WO1997041208A1 (en) | 1996-04-26 | 1997-11-06 | Case Western Reserve University | Skin regeneration using mesenchymal stem cells |
US5827740A (en) | 1996-07-30 | 1998-10-27 | Osiris Therapeutics, Inc. | Adipogenic differentiation of human mesenchymal stem cells |
AU5436998A (en) | 1996-11-15 | 1998-06-03 | Osiris Therapeutics, Inc. | MSC-megakaryocyte precursor composition and method of isolating MSCs asso ciated with isolated megakaryocytes by isolating megakaryocytes |
JP4531135B2 (en) | 1996-12-06 | 2010-08-25 | オシリス セラピューティクス,インコーポレイテッド | Improved chondrogenic differentiation of human mesenchymal stem cells |
WO1998051317A1 (en) | 1997-05-13 | 1998-11-19 | Osiris Therapeutics, Inc. | Osteoarthritis cartilage regeneration using human mesenchymal stem cells |
US5786207A (en) * | 1997-05-28 | 1998-07-28 | University Of Pittsburgh | Tissue dissociating system and method |
EP1028737B1 (en) | 1997-07-03 | 2007-04-04 | Osiris Therapeutics, Inc. | Human mesenchymal stem cells from peripheral blood |
ES2251773T5 (en) | 1997-07-14 | 2009-05-12 | Osiris Therapeutics, Inc. | REGENERATION OF THE CARDIAC MUSCLE USING MESENQUIMATOSE PRECURSOR CELLS. |
US6077987A (en) | 1997-09-04 | 2000-06-20 | North Shore-Long Island Jewish Research Institute | Genetic engineering of cells to enhance healing and tissue regeneration |
PT1007559E (en) * | 1997-09-05 | 2006-05-31 | Yissum Res Dev Co | GENETICALLY MODIFIED CELLS EXPRESSING BONE MORPHOGENETIC PROTEINS |
FR2771102B1 (en) * | 1997-11-14 | 2000-12-15 | Biocoral Inc | METHOD FOR INDUCING OSTEOBLASTIC DIFFERENTIATION OF HUMAN EXTRAMEDULAR FAT TISSUE CELLS |
CA2312847C (en) * | 1997-12-02 | 2014-09-30 | Zen Bio, Inc. | Differentiation of adipose stromal cells into osteoblasts and uses thereof |
US6866842B1 (en) * | 1998-05-01 | 2005-03-15 | University Of Pittsburgh | Muscle-derived cells (MDCs) for treating muscle-or bone-related injury or dysfunction |
US6153432A (en) * | 1999-01-29 | 2000-11-28 | Zen-Bio, Inc | Methods for the differentiation of human preadipocytes into adipocytes |
US6777231B1 (en) * | 1999-03-10 | 2004-08-17 | The Regents Of The University Of California | Adipose-derived stem cells and lattices |
KR100968165B1 (en) | 1999-03-10 | 2010-07-06 | 더 리전츠 오브 더 유니버시티 오브 캘리포니아 | Adipose-derived stem cells and lattices |
US20050076396A1 (en) * | 1999-03-10 | 2005-04-07 | Katz Adam J. | Adipose-derived stem cells and lattices |
US6429013B1 (en) * | 1999-08-19 | 2002-08-06 | Artecel Science, Inc. | Use of adipose tissue-derived stromal cells for chondrocyte differentiation and cartilage repair |
US7078232B2 (en) * | 1999-08-19 | 2006-07-18 | Artecel, Inc. | Adipose tissue-derived adult stem or stromal cells for the repair of articular cartilage fractures and uses thereof |
US6555374B1 (en) * | 1999-08-19 | 2003-04-29 | Artecel Sciences, Inc. | Multiple mesodermal lineage differentiation potentials for adipose tissue-derived stromal cells and uses thereof |
AU3869501A (en) | 2000-02-26 | 2001-09-03 | Artecel Sciences Inc | Pleuripotent stem cells generated from adipose tissue-derived stromal cells and uses thereof |
US7311905B2 (en) * | 2002-02-13 | 2007-12-25 | Anthrogenesis Corporation | Embryonic-like stem cells derived from post-partum mammalian placenta, and uses and methods of treatment using said cells |
JP2005508393A (en) * | 2001-11-09 | 2005-03-31 | アーテセル・サイエンシズ・インコーポレーテツド | Methods and compositions for the use of stromal cells to support embryonic and adult stem cells |
PL370261A1 (en) | 2001-11-09 | 2005-05-16 | Artecel Sciences, Inc. | Adipose tissue-derived stromal cells for the repair of corneal and intra-orbital defects and uses thereof |
US7020590B2 (en) * | 2001-12-28 | 2006-03-28 | Intel Corporation | Simulation of di/dt-induced power supply voltage variation |
WO2003057152A2 (en) * | 2001-12-31 | 2003-07-17 | Isolagen Technologies, Inc | Hair follicle growth |
WO2003084468A2 (en) * | 2002-04-03 | 2003-10-16 | Artecel Sciences, Inc. | Improvements of adipocytic differentiated adipose derived adult stem cells and uses thereof |
EP1527161B1 (en) * | 2002-07-31 | 2015-10-28 | Yves Saint-Laurent Parfums | Stem cells derived from adipous tissue and differentiated cells derived from said cells |
-
2005
- 2005-07-29 US US11/192,753 patent/US7531355B2/en active Active
-
2006
- 2006-07-18 WO PCT/US2006/028040 patent/WO2007015906A2/en active Application Filing
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4443546A (en) * | 1980-07-07 | 1984-04-17 | The Beth Israel Hospital Association | Process and composition for propagating mammalian cells |
US5605687A (en) * | 1992-05-15 | 1997-02-25 | Arch Development Corporation | Methods and compositions of a polymer (poloxamer) for repair of electrical injury |
US20030161816A1 (en) * | 2001-12-07 | 2003-08-28 | Fraser John K. | Systems and methods for treating patients with processed lipoaspirate cells |
US20040193274A1 (en) * | 2003-03-28 | 2004-09-30 | Trieu Hai H. | Materials and methods for augmenting and/or repairing intervertebral discs |
Non-Patent Citations (1)
Title |
---|
CHARBORD, P. ET AL.: "Analysis of the microenvironment necessary for engraftment: role of the vascular smooth muscle-like stromal cells.", J HEMATOTHERAPY STEM CELL RES., vol. 9, 2000, pages 935 - 943 * |
Also Published As
Publication number | Publication date |
---|---|
WO2007015906A2 (en) | 2007-02-08 |
US7531355B2 (en) | 2009-05-12 |
US20070025972A1 (en) | 2007-02-01 |
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