CN100372928C - 灵长类动物胚胎干细胞的无血清培养物 - Google Patents

灵长类动物胚胎干细胞的无血清培养物 Download PDF

Info

Publication number
CN100372928C
CN100372928C CNB018062350A CN01806235A CN100372928C CN 100372928 C CN100372928 C CN 100372928C CN B018062350 A CNB018062350 A CN B018062350A CN 01806235 A CN01806235 A CN 01806235A CN 100372928 C CN100372928 C CN 100372928C
Authority
CN
China
Prior art keywords
embryonic stem
stem cells
serum
substratum
growth factor
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 - Lifetime
Application number
CNB018062350A
Other languages
English (en)
Other versions
CN1416345A (zh
Inventor
J·A·汤姆森
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Wisconsin Alumni Research Foundation
Original Assignee
Wisconsin Alumni Research Foundation
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Family has litigation
First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=24079156&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=CN100372928(C) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Application filed by Wisconsin Alumni Research Foundation filed Critical Wisconsin Alumni Research Foundation
Publication of CN1416345A publication Critical patent/CN1416345A/zh
Application granted granted Critical
Publication of CN100372928C publication Critical patent/CN100372928C/zh
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Classifications

    • 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
    • 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
    • C12N5/0603—Embryonic cells ; Embryoid bodies
    • C12N5/0606—Pluripotent embryonic cells, e.g. embryonic stem cells [ES]
    • 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
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/90—Serum-free medium, which may still contain naturally-sourced components
    • 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
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/10—Growth factors
    • C12N2501/115—Basic fibroblast growth factor (bFGF, FGF-2)

Abstract

本文公开了培养灵长类动物胚胎干细胞的方法。在长时间和稳定的基础上，在外源提供的成纤维细胞生长因子的存在下和缺乏动物血清的条件下培养这些细胞。较佳的是，培养基中还存在成纤维细胞饲养层。本发明还公开了含有成纤维细胞饲养层和成纤维细胞生长因子的培养基。

Description

灵长类动物胚胎干细胞的无血清培养物
相关申请的交叉参考
不适用。
关于联邦政府资助的研究的申明
发明的背景
本发明涉及培养灵长类动物胚胎干细胞培养物的方法和培养中使用到的培养基。
已从植入前胚胎获得灵长类动物(如猴和人)多能胚胎干细胞。参见美国专利第5,843,780号；和J.Thomson等人，282Science，1145-1147(1998)。这些出版物和所有其它本文涉及的出版物的公开内容都被完整地纳入本文作为参考。尽管经过长时间的培养，但这些细胞仍稳定地维持着发育成为所有三种胚胎性胚层的高级衍生物的潜力。
已在人发育生物学、药物开发、药物测试和移植医学中广泛使用灵长类动物(具体是人)的胚胎干细胞系。例如，目前关于植入后人胚胎的知识在很大程度上以有限量的静态组织切片为基础。考虑到伦理方面的因素，人们基本上仍未对控制人胚胎早期的发育选择的基础机制进行研究。
虽然小鼠是实验哺乳动物发育生物学的主要研究对象，并且小鼠和人之间许多控制发育的基本机制是保守的，但是在小鼠和人的早期发育中仍存在明显的差异。因此，灵长类动物/人ES细胞应可提供关于它们的分化和功能的新的重要视点。
可使用灵长类动物ES细胞的分化衍生物来鉴别新药的基因靶标、测试新化合物的毒性或致畸性以及用于移植以替换疾病细胞群。可通过移植ES细胞衍生的细胞来进行治疗的潜在病症包括帕金森综合症、心肌梗塞、青少年发作的糖尿病和白血病。例如可参见J.Rossant等人，17Nature Biotechnology，23-4(1999)和J.Gearhart，282Sciencc，1061-2(1998)。
关于灵长类动物的胚胎干细胞培养物的利用，其关键因素是长期的增殖能力、长时间培养后的发育潜力和核型稳定性。这些细胞的培养物(尤其是培养在成纤维细胞饲养层上的培养物)一般补加动物血清(尤其是胎牛血清)，以便在这样的培养期间产生所需的增殖。
例如，在美国专利第5,453,357、5,670,372和5,690,296中描述了各种培养条件，包括一些同时使用一类碱性成纤维细胞生长因子和动物血清的条件。不幸的是，不同批次的血清倾向于具有不同的特性，从而影响了培养特征。
在WO98/30679中讨论了使用无血清添加剂类替换动物血清，以支持培养中某些胚胎干细胞的生长。该血清替代物包括清蛋白或清蛋白替代品、一种或多种氨基酸、一种或多种维生素、一种或多种运铁蛋白或运铁蛋白的替代品、一种或多种抗氧化剂、一种或多种胰岛素或胰岛素替代品、一种或多种胶原蛋白前体以及一种或多种痕量元素。应注意的是，还可在这种替代物中添加白血病抑制因子、青灰因子或睫状神经营养因子。不幸的是，在关于灵长类动物的胚胎干细胞培养物(尤其是生长在成纤维细胞饲养层上的那些培养物)的内容中，这些培养基并未证明是让人满意的。
在关于营养血清培养基(如胎牛血清)的内容中，WO99/20741讨论了在培养灵长类动物的干细胞中使用各种生长因子(如bFGF)的益处。但是，文中并未描述到不含有营养血清的培养基。
美国专利第5,405,772号描述了用于造血细胞和骨髓基质细胞的生长培养基。为了达到目的，文中建议在无血清的培养基中使用成纤维细胞生长因子。但是，文中并未描述灵长类动物胚胎干细胞的生长条件。
由此可见，还需要不使用动物血清即可稳定地培养灵长类胚胎干细胞的技术。
发明概要
一方面，本发明提供一种培养灵长类动物的胚胎干细胞的方法。在基本上没有哺乳动物胎儿血清(较佳还基本上没有任何动物血清)的培养基中、在成纤维细胞生长因子的存在下培养干细胞，其中，所述生长因子的来源不仅仅是成纤维细胞饲养层。在较佳的实施方式中，该培养基还具有成纤维细胞饲养层。
成纤维细胞生长因子是哺乳动物发育的主要分子。目前有9种已知的成纤维细胞生长因子配体和4种针对它们的信号传导的成纤维细胞生长因子受体(以及它们的剪接变体)。综述可参见D.Ornitz等人，25 J.Biol.Chem.，15292-7(1996)；美国专利第5,453,357号。在不同物种间这些因子会存在细微的差别，因此术语成纤维细胞生长因子不局限于物种。但是，我优选使用人的成纤维细胞生长因子，更优选使用由重组基因产生的人的碱性成纤维细胞生长因子。易于从Gibco BRL-Life Techologies和其它公司获得大量的这种化合物。
应注意的是，为了本专利的目的，所述培养基还需要基本上不含有特定的血清，即使已经从血清中分离出分散的成分(如牛血清清蛋白)并在随后以外来的方式补充。问题在于，当加入血清本身时，差异性问题就会产生。但是，当加入血清的一种或多种已鉴别清楚的纯化成分时，它们就产生差异性的问题。
较佳的是，采用本发明方法培养的灵长类动物胚胎干细胞是人的胚胎干细胞，它们是真正的ES细胞系，因为它们能(i)以未分化的状态在体外无限增殖；(ii)即使在长时间培养后仍可分化成所有3个胚胎性胚层(内胚层、中胚层和外胚层)的衍生物；(iii)在整个长时间的培养过程中维持正常核型。因此，它们是多能的。
上述培养使所述胚胎干细胞可在培养基中稳定地增殖1个月以上(较佳6个月以上，更佳12个月以上)，而同时使所述干细胞维持分化成内胚层、中胚层和外胚层组织的衍生物的潜力，以及维持它们的核型。
另一方面，本发明提供培养灵长类动物胚胎干细胞的另一种方法。在基本上不含有哺乳动物胎儿血清(较佳还基本上不含有任何动物的血清)的培养基中、在能激活成纤维细胞生长因子信号受体的生长因子的存在下培养干细胞，其中，提供所述生长因子的来源不仅仅是成纤维细胞饲养层。虽然所述生长因子较佳是成纤维细胞生长因子，但是它也可以是其它物质，如设计用来激活成纤维细胞生长因子受体的某些合成的小肽(如由重组DNA变体或突变体产生的肽)。综述可参见T.Yamaguchi等人，152Dev.Biol.，75-88(1992)(信号受体)。
又一方面，本发明提供用于培养灵长类动物胚胎干细胞的培养系统。该系统具有成纤维细胞饲养层和人碱性成纤维细胞生长因子，该生长因子的来源不仅仅是该成纤维细胞饲养层。该培养系统基本上不含有动物血清。
又一方面，本发明提供采用上述方法衍生获得的细胞系(较佳是克隆的细胞系)。“衍生”在此使用其广义含义，覆盖了直接或间接衍生的细胞系。
因而，本发明避免了由于动物血清批次差异而产生的差异性。此外，已发现在避免使用动物血清的同时，使用成纤维细胞生长因子可增加克隆的效率。
因此，本发明的一个优点是提供用于灵长类动物胚胎干细胞系的培养条件，其中，所述条件变化少，并使得克隆更有效率。在研究了本说明书和权利要求书后，本发明的其它优点将是显而易见的。
优选实施方式的详细描述
在下述实验中，我使用了本发明的方法和培养系统来培养人ES细胞系。两个克隆衍生的人ES细胞系在克隆产生后增殖了8个月以上，并且维持了分化成所有3个胚胎性胚层的高级衍生物的能力。
人ES细胞系H9的初始衍生、培养和表征的技术在J.Thomson等人(282Science，1145-1147(1998))的文章中有所描述。在我的实验中，在完成上述操作后，在经辐射的(35戈瑞的γ辐射过的)小鼠胚胎成纤维细胞上平板培养人ES细胞。此步骤中使用的培养基由80％“KnockOut”Dulbeco改进的Eagle培养基(DMEM)(Gibco BRL，Rockville，MD)、1mM L-谷氨酰胺、0.1mM β-巯基乙醇和1％非必需氨基酸原液(Gibco BRL Rockville，MD)组成，补充了20％胎牛血清(HyCloneLogan，UT)或20％KnockOut SR(最初优化用于小鼠ES细胞的无血清替代品，Gibco BRL，Rockville，MD)。KnockOut SR的成分是WO98/30679中描述的用于血清替代品的成分。
在另一实验中，培养基中补加血清或者前述血清替代品KnockOut SR，还可含有或不含人重组碱性成纤维细胞生长因子(bFGF，4ng/m1)。培养基中bFGF的浓度范围较佳是0.1ng/m1到500ng/m1之间。
为了测定在不同培养条件下的克隆率，使用0.05％胰蛋白酶/0.25％EDTA，处理7分钟，将H-9培养物解离成单个细胞，然后离心洗涤，之后放到有丝分裂失活的小鼠胚胎成纤维细胞上培养(105个ES细胞/孔，6孔平板)。为了证实单个细胞的生长是由克隆的ES细胞系衍生的，在立体显微镜的直接观察下挑选出各个细胞，然后使用微移液管将其转移到含有小鼠成纤维细胞饲养层以及含有20％血清替代品和4ng/ml bFGF的培养基的96孔平板的各个孔中。
通过每5-7天使用1mg/m1的胶原蛋白酶IV型(Gibco BRL，Rockville，MD)，进行常规的传代培养，使这些克隆得到增殖。衍生6个月后，使用标准的G-显带技术(分析20条染色体的伸展情况)测得H9细胞具有正常的XX核型。但是，衍生后7个月，在单个核型制品中，20条染色体伸展中有16条具有正常的XX核型，而有4条伸展显示出随机的异常性，其中包括1条具有向染色体13短臂的移位，1条具有反向的染色体20，1条具有向染色体4的短臂上的移位，1条具有多个碎片。结果，在衍生8、10和12.75个月后，H9细胞在所检测的所有20条染色体伸展中具有正常的核型。
我们观察到，在先前所述的包括动物血清的培养条件下人ES细胞的克隆率差(不论存在bFGF与否)。我们还观察到，在缺乏动物血清时，克隆率增加，并且存在bFGF时效率增加得更高。
下面的数据是由平板培养的105个单独的ES细胞产生的集落总数+/-平均值的标准误差(集落克隆率％)。含有20％胎牛血清但没有bFGF的培养结果是240±28个。含有20％血清并含有bFGF的培养结果大约相同，为260±12个。缺乏血清(存在20％血清替代品)也没有bFGF所得的结果是633±43个，但含有bFGF的结果是826±61个。因此，血清对克隆率产生不利的影响，存在bFGF而不含有血清的条件对克隆率产生了协同的利益。
在存在血清的条件下长时间培养人ES细胞不需要从外源提供bFGF，并且向含有血清的培养基中加入bFGF也没有明显地提高人ES细胞克隆率(如上所述)。但是，在没有血清的培养基中，bFGF提高了人ES细胞的初始克隆率。
此外，我还发现，对于灵长类动物胚胎干细胞在无动物血清的条件下的连续的未分化繁殖来说，提供外源bFGF是非常重要的。在缺乏bFGF的无血清培养基中，在培养两周后，人ES细胞一律进行分化。(在缺乏bFGF的条件下)其它因子(如LIF)的加入并未能阻止这种分化。
所察觉到的结果特别适用于克隆系。在这方面，在直接的显微镜观察的帮助下，通过将细胞单独放到96孔平板的孔中，可筛选出用于扩增的克隆。在放到96孔平板的孔中培养的192个H-9细胞中，两个克隆成功地扩增(H-9.1和H-9.2)。接着在补充了血清替代品和bFGF的培养基中连续培养这两个克隆。
即使在克隆后连续培养了8个月以上，H9.1和H9.2细胞仍都维持了正常的XX核型。所述H9.1和H9.2克隆即使在无血清培养基中长时间培养后，仍维持形成所有三个胚胎性胚层的衍生物的潜力。在培养6个月后，H9.1和H9.2克隆被证实具有正常的核型，然后用它们给SCID-米色小鼠注射。
H9.1和H9.2细胞都形成了含有所有三个胚胎性胚层的衍生物的畸胎瘤，包括肠上皮(内胚层)，胚肾、横纹肌、平滑肌、骨、软骨(中胚层)和神经组织(外胚层)。H9.1和H9.2细胞的高传代细胞的畸胎瘤中所观察到的分化范围，与低传代的H9亲代细胞形成的畸胎瘤内的分化范围是相当的。
从上面所述内容应可理解，虽然动物血清对生长起支持性作用，但是它是一种复杂的混合物，它可同时含有对人ES细胞培养有利或有害的化合物。而且，不同批次的血清支持人ES细胞有力地以未分化状态增殖的能力极大地不同。使用清楚确定的成分替换血清的做法减少了与这种血清批次的变化有关的结果的差异性，并能进行更精细的分化研究。
此外，含有血清的培养基中的较低的克隆率表明，在常规使用的血清中存在对干细胞生存不利的化合物，尤其是当细胞被分散成单个细胞时。因此非常需要避免使用这些化合物。
上文已结合优选实施方式对本发明进行了描述。这一原理的其它形式也打算包括在权利要求所述的范围内。例如，虽然在上述实验中使用了重组产生的人碱性成纤维细胞生长因子，但是天然分离获得的成纤维细胞生长因子也适合使用。此外，这些技术也应证明可适用于猴和其它灵长类动物的细胞培养物。
因此，为判断本发明的整个范围，应查阅权利要求书。
工业应用性
本发明提供用于培养灵长类动物胚胎干细胞的方法，和培养过程中使用到的培养基。

Claims (15)

1.一种培养灵长类动物胚胎干细胞的方法，它包括：
在不含哺乳动物胎儿血清而含有清蛋白或清蛋白替代品、一种或多种氨基酸、一种或多种维生素、一种或多种运铁蛋白或运铁蛋白的替代品、一种或多种抗氧化剂、一种或多种胰岛素或胰岛素替代品、一种或多种胶原蛋白前体以及一种或多种痕量元素的培养基中、在成纤维细胞生长因子的存在下培养所述干细胞，其中，所述生长因子的来源不仅仅是成纤维细胞饲养层，所述细胞是非人灵长类动物胚胎干细胞。
2.如权利要求1所述的方法，其特征在于，所述培养基不含有任何动物血清。
3.如权利要求2所述的方法，其特征在于，所述培养基还含有成纤维细胞饲养层。
4.如权利要求2所述的方法，其特征在于，所述成纤维细胞生长因子是碱性成纤维细胞生长因子。
5.如权利要求4所述的方法，其特征在于，所述成纤维细胞生长因子是已由重组基因产生的人碱性成纤维细胞生长因子。
6.如权利要求2所述的方法，其特征在于，所述方法包括，使所述胚胎干细胞在所述培养基中增殖1个月以上。
7.如权利要求5所述的方法，其特征在于，在所述方法的至少一部分中，所述人碱性成纤维细胞生长因子的浓度至少为0.1ng/ml。
8.如权利要求1所述的方法，其特征在于，所述培养基补充有血清替代品KnockOut SR。
9.一种培养灵长类动物胚胎干细胞的方法，它包括：
在不含有哺乳动物胎儿血清而含有清蛋白或清蛋白替代品、一种或多种氨基酸、一种或多种维生素、一种或多种运铁蛋白或运铁蛋白的替代品、一种或多种抗氧化剂、一种或多种胰岛素或胰岛素替代品、一种或多种胶原蛋白前体以及一种或多种痕量元素的培养基中、在能激活成纤维细胞生长因子信号受体的生长因子的存在下培养干细胞，其中，所述能激活信号受体的生长因子的来源不仅仅是成纤维细胞饲养层，所述细胞是非人灵长类动物胚胎干细胞。
10.如权利要求9所述的方法，其特征在于，所述培养基不含任何动物血清。
11.如权利要求10所述的方法，其特征在于，所述培养基还含有成纤维细胞饲养层。
12.如权利要求10所述的方法，其特征在于，所述方法包括，使所述胚胎干细胞在所述培养基中增殖1个月以上。
13.一种用于培养灵长类动物胚胎干细胞的培养系统，它含有：
成纤维细胞生长因子；和
清蛋白或清蛋白替代品、一种或多种氨基酸、一种或多种维生素、一种或多种运铁蛋白或运铁蛋白的替代品、一种或多种抗氧化剂、一种或多种胰岛素或胰岛素替代品、一种或多种胶原蛋白前体以及一种或多种痕量元素；
其中，所述培养系统不含动物血清。
14.如权利要求13所述的培养系统，其特征在于，所述培养系统还含有成纤维细胞饲养层，且其中所述成纤维细胞生长因子的来源不仅仅是该成纤维细胞饲养层。
15.权利要求13或14所述的培养系统在培养灵长类动物胚胎干细胞中的用途。
CNB018062350A 2000-03-09 2001-03-02 灵长类动物胚胎干细胞的无血清培养物 Expired - Lifetime CN100372928C (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US09/522,030 US7005252B1 (en) 2000-03-09 2000-03-09 Serum free cultivation of primate embryonic stem cells
US09/522,030 2000-03-09

Publications (2)

Publication Number Publication Date
CN1416345A CN1416345A (zh) 2003-05-07
CN100372928C true CN100372928C (zh) 2008-03-05

Family

ID=24079156

Family Applications (1)

Application Number Title Priority Date Filing Date
CNB018062350A Expired - Lifetime CN100372928C (zh) 2000-03-09 2001-03-02 灵长类动物胚胎干细胞的无血清培养物

Country Status (15)

Country Link
US (4) US7005252B1 (zh)
EP (1) EP1261691B1 (zh)
JP (3) JP5717311B2 (zh)
KR (1) KR100795760B1 (zh)
CN (1) CN100372928C (zh)
AU (2) AU2001241973B2 (zh)
BR (1) BR0108507A (zh)
CA (1) CA2402299C (zh)
HK (1) HK1053616A1 (zh)
IL (2) IL151270A0 (zh)
IS (1) IS6515A (zh)
MX (1) MXPA02008698A (zh)
NO (1) NO335780B1 (zh)
NZ (1) NZ520701A (zh)
WO (1) WO2001066697A2 (zh)

Families Citing this family (151)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7410798B2 (en) 2001-01-10 2008-08-12 Geron Corporation Culture system for rapid expansion of human embryonic stem cells
US7413904B2 (en) * 1998-10-23 2008-08-19 Geron Corporation Human embryonic stem cells having genetic modifications
US10638734B2 (en) 2004-01-05 2020-05-05 Abt Holding Company Multipotent adult stem cells, sources thereof, methods of obtaining and maintaining same, methods of differentiation thereof, methods of use thereof and cells derived thereof
US8252280B1 (en) 1999-08-05 2012-08-28 Regents Of The University Of Minnesota MAPC generation of muscle
US7015037B1 (en) 1999-08-05 2006-03-21 Regents Of The University Of Minnesota Multiponent adult stem cells and methods for isolation
US7838289B2 (en) 2001-02-14 2010-11-23 Abt Holding Company Assay utilizing multipotent adult stem cells
US7455983B2 (en) * 2000-01-11 2008-11-25 Geron Corporation Medium for growing human embryonic stem cells
US20100173410A1 (en) * 2000-03-09 2010-07-08 Wicell Research Institute, Inc. Cultivation of Primate Embryonic Stem Cells
US7439064B2 (en) * 2000-03-09 2008-10-21 Wicell Research Institute, Inc. Cultivation of human embryonic stem cells in the absence of feeder cells or without conditioned medium
WO2003008535A2 (en) * 2001-07-20 2003-01-30 Technion Research And Development Foundation Ltd. Methods of generating human cardiac cells and tissues and uses thereof
WO2003046129A2 (en) * 2001-11-09 2003-06-05 Trustees Of The University Of Pennsylvania Compositions for the derivation of germ cells from stem cells and methods of use thereof
GB0202149D0 (en) 2002-01-30 2002-03-20 Univ Edinburgh Pluripotency determining factors and uses thereof
US20040111285A1 (en) * 2002-04-09 2004-06-10 Mark Germain Method for human pluripotent stem cells
AU2003260724A1 (en) * 2002-08-22 2004-03-11 Celltran Limited Cell culture surface
AU2002344527A1 (en) * 2002-10-25 2004-05-13 Hunan Hui-Lin Life Technology Co. The feeder cell layer for in vitro culturing human embryonic stem cells and the method for culturing embryonic stem cells
GB0304918D0 (en) * 2003-03-05 2003-04-09 Celltran Ltd Cell culture
US7820439B2 (en) 2003-09-03 2010-10-26 Reliance Life Sciences Pvt Ltd. In vitro generation of GABAergic neurons from pluripotent stem cells
US20070269412A1 (en) 2003-12-02 2007-11-22 Celavie Biosciences, Llc Pluripotent cells
WO2005056755A2 (en) 2003-12-02 2005-06-23 Catholic Healthcare West Compositions and methods for propagation of neural progenitor cells
US7985585B2 (en) 2004-07-09 2011-07-26 Viacyte, Inc. Preprimitive streak and mesendoderm cells
US20050266554A1 (en) * 2004-04-27 2005-12-01 D Amour Kevin A PDX1 expressing endoderm
EP1709159B1 (en) 2003-12-23 2019-05-15 Viacyte, Inc. Definitive endoderm
US7541185B2 (en) * 2003-12-23 2009-06-02 Cythera, Inc. Methods for identifying factors for differentiating definitive endoderm
US8647873B2 (en) 2004-04-27 2014-02-11 Viacyte, Inc. PDX1 expressing endoderm
US7625753B2 (en) * 2003-12-23 2009-12-01 Cythera, Inc. Expansion of definitive endoderm cells
US8586357B2 (en) * 2003-12-23 2013-11-19 Viacyte, Inc. Markers of definitive endoderm
US20050233446A1 (en) * 2003-12-31 2005-10-20 Parsons Xuejun H Defined media for stem cell culture
US20080085554A1 (en) * 2004-02-13 2008-04-10 Norio Nakatsuji Culture Medium for Culturing Feeder Cells for Embryonic Stem Cells Culture and the Prepared Feeder Cells
SG150567A1 (en) 2004-03-23 2009-03-30 Akaike Toshihiro Pluripotent stem cell growing method
EP2377922B1 (en) 2004-04-27 2020-04-08 Viacyte, Inc. PDX1 expressing endoderm
EP1749091B1 (en) * 2004-05-21 2013-07-31 Wicell Research Institute, Inc. Feeder independent extended culture of embryonic stem cells
CA2573354C (en) 2004-07-09 2015-02-24 Cythera, Inc. Preprimitive streak and mesendoderm cells
MX2007001772A (es) * 2004-08-13 2007-07-11 Univ Georgia Res Found Composiciones y metodos para auto-renovacion y diferenciacion de celulas troncales embrionicas humanas.
CN101044235B (zh) 2004-09-08 2013-01-02 威斯康星校友研究基金会 胚胎干细胞的培养基和培养
ES2800973T3 (es) * 2004-09-28 2021-01-07 Wisconsin Alumni Res Found Cultivo de células madre embrionarias de primates
US8017395B2 (en) 2004-12-17 2011-09-13 Lifescan, Inc. Seeding cells on porous supports
WO2006073966A1 (en) * 2004-12-30 2006-07-13 Stemlifeline, Inc. Methods and systems relating to embryonic stem cell lines
US20060275899A1 (en) * 2004-12-30 2006-12-07 Stemlifeline, Inc. Methods and compositions relating to embryonic stem cell lines
CN1298843C (zh) * 2005-02-07 2007-02-07 十堰市太和医院 不依赖饲养细胞的人胚干细胞培养基
AU2006202209B2 (en) * 2005-05-27 2011-04-14 Lifescan, Inc. Amniotic fluid derived cells
WO2006133052A2 (en) * 2005-06-08 2006-12-14 Centocor, Inc. A cellular therapy for ocular degeneration
WO2007002086A2 (en) * 2005-06-22 2007-01-04 Geron Corporation Suspension culture of human embryonic stem cells
WO2007016366A2 (en) * 2005-07-29 2007-02-08 Yale University Defined culture conditions of human embryonic stem cells
KR100670616B1 (ko) * 2005-08-25 2007-01-17 주식회사 메디아나전자 상피세포 또는 섬유아세포를 이용한 세포 배양용 인공 배아 및 세포 배양 장치
AU2006285467A1 (en) * 2005-09-02 2007-03-08 Agency For Science, Technology And Research Method of deriving mesenchymal stem cells
WO2007044572A2 (en) * 2005-10-06 2007-04-19 University Of Massachusetts Cell division marker
AU2006301549B2 (en) * 2005-10-07 2013-02-14 Cellartis Ab A method for obtaining a xeno-free hBS cell line
ES2687233T3 (es) 2005-10-27 2018-10-24 Viacyte, Inc. Endodermo de intestino proximal dorsal y ventral que expresa PDX-1
US7413900B2 (en) * 2005-10-31 2008-08-19 President And Fellows Of Harvard College Immortalized fibroblasts
US7695965B2 (en) 2006-03-02 2010-04-13 Cythera, Inc. Methods of producing pancreatic hormones
US11254916B2 (en) 2006-03-02 2022-02-22 Viacyte, Inc. Methods of making and using PDX1-positive pancreatic endoderm cells
SG10201405380QA (en) 2006-03-02 2014-10-30 Cythera Inc Endocrine precursor cells, pancreatic hormone-expressing cells and methods of production
US7989204B2 (en) 2006-04-28 2011-08-02 Viacyte, Inc. Hepatocyte lineage cells
EP2021462B1 (en) 2006-04-28 2019-01-09 Lifescan, Inc. Differentiation of human embryonic stem cells
US8741643B2 (en) 2006-04-28 2014-06-03 Lifescan, Inc. Differentiation of pluripotent stem cells to definitive endoderm lineage
CN100465268C (zh) * 2006-05-17 2009-03-04 北京大学 人胚胎干细胞的培养方法及其专用培养基
US7964402B2 (en) 2006-05-25 2011-06-21 Sanford-Burnham Medical Research Institute Methods for culture and production of single cell populations of human embryonic stem cells
JP2009540826A (ja) * 2006-06-20 2009-11-26 ジェンザイム・コーポレーション 軟骨細胞増幅のための無血清培地およびその使用
KR20090085019A (ko) * 2006-08-11 2009-08-06 주식회사 엠씨티티 줄기세포 및 피더세포의 공배양용 배지 조성물
US20100323442A1 (en) * 2006-10-17 2010-12-23 Emmanuel Edward Baetge Modulation of the phosphatidylinositol-3-kinase pathway in the differentiation of human embryonic stem cells
AU2007321928A1 (en) 2006-11-24 2008-05-29 Regents Of The University Of Minnesota Endodermal progenitor cells
US10829733B2 (en) * 2007-01-04 2020-11-10 Biolamina Ab Composition and method for enabling proliferation of pluripotent human stem cells
US7883698B2 (en) * 2007-01-17 2011-02-08 Maria Michejda Isolation and preservation of fetal hematopoietic and mesencymal system cells from non-controversial materials and/or tissues resulting from miscarriages and methods of therapeutic use
KR20090115142A (ko) * 2007-01-30 2009-11-04 유니버시티 오브 조지아 리서치 파운데이션, 인코포레이티드 초기 중배엽 세포，내배엽 및 중배엽 계통의 생성에 유용한 중내배엽 세포의 안정한 집단 및 다능성 유주 세포（ｍｍｃ）
US7951593B2 (en) * 2007-03-20 2011-05-31 Universite Rene Descartes-Paris V Culture medium for gingival fibroblasts
KR101732952B1 (ko) 2007-07-01 2017-05-08 라이프스캔, 인코포레이티드 단일 다분화성 줄기 세포 배양
US9080145B2 (en) 2007-07-01 2015-07-14 Lifescan Corporation Single pluripotent stem cell culture
CN101861386A (zh) 2007-07-18 2010-10-13 生命扫描有限公司 人胚胎干细胞的分化
DK2185693T3 (da) 2007-07-31 2019-09-23 Lifescan Inc Differentiering af humane embryoniske stamceller
ES2665434T3 (es) 2007-07-31 2018-04-25 Lifescan, Inc. Diferenciación de células madre pluripotentes usando células alimentadoras humanas
US7695963B2 (en) 2007-09-24 2010-04-13 Cythera, Inc. Methods for increasing definitive endoderm production
CN107574142B (zh) * 2007-11-27 2021-07-06 生命扫描有限公司 人胚胎干细胞的分化
US20100087002A1 (en) * 2008-02-21 2010-04-08 Benjamin Fryer Methods, Surface Modified Plates and Compositions for Cell Attachment, Cultivation and Detachment
CA2715878C (en) 2008-02-21 2017-06-13 Centocor Ortho Biotech Inc. Methods, surface modified plates and compositions for cell attachment, cultivation and detachment
WO2009131568A1 (en) 2008-04-21 2009-10-29 Cythera, Inc. Methods for purifying endoderm and pancreatic endoderm cells derived from human embryonic stem cells
US8338170B2 (en) 2008-04-21 2012-12-25 Viacyte, Inc. Methods for purifying endoderm and pancreatic endoderm cells derived from human embryonic stem cells
US7939322B2 (en) 2008-04-24 2011-05-10 Centocor Ortho Biotech Inc. Cells expressing pluripotency markers and expressing markers characteristic of the definitive endoderm
US8623648B2 (en) * 2008-04-24 2014-01-07 Janssen Biotech, Inc. Treatment of pluripotent cells
EP3851520A1 (en) 2008-04-30 2021-07-21 SanBio, Inc. Neural regenerating cells with alterations in dna methylation
US20090298178A1 (en) * 2008-06-03 2009-12-03 D Amour Kevin Allen Growth factors for production of definitive endoderm
DK2297319T3 (en) 2008-06-03 2015-10-19 Viacyte Inc GROWTH FACTORS FOR PREPARING DEFINITIVE ENDODERM
KR20180018839A (ko) 2008-06-30 2018-02-21 얀센 바이오테크 인코포레이티드 만능 줄기 세포의 분화
WO2010002785A1 (en) * 2008-06-30 2010-01-07 Centocor Ortho Biotech Inc. Differentiation of pluripotent stem cells
CA2731821A1 (en) 2008-07-25 2010-01-28 Stephen Dalton Compositions for mesoderm derived isl1+ multipotent cells (imps), epicardial progenitor cells (epcs) and multipotent cxcr4+cd56+ cells (c56cs) and methods of use
US20110305672A1 (en) 2008-07-25 2011-12-15 University Of Georgia Research Foundation, Inc. COMPOSITIONS FOR MESODERM DERIVED ISL1+ MULTIPOTENT CELLS (IMPs), EPICARDIAL PROGENITOR CELLS (EPCs) AND MULTIPOTENT CD56C CELLS (C56Cs) AND METHODS OF PRODUCING AND USING SAME
US20100028307A1 (en) * 2008-07-31 2010-02-04 O'neil John J Pluripotent stem cell differentiation
JP2012507289A (ja) 2008-10-31 2012-03-29 ヤンセン バイオテツク，インコーポレーテツド ヒト胚性幹細胞の膵内分泌系への分化
US9234178B2 (en) 2008-10-31 2016-01-12 Janssen Biotech, Inc. Differentiation of human pluripotent stem cells
CA2907326A1 (en) 2008-11-04 2010-05-14 Chad Green Stem cell aggregate suspension compositions and methods for differentiation thereof
EP2356227B1 (en) 2008-11-14 2018-03-28 Viacyte, Inc. Encapsulation of pancreatic cells derived from human pluripotent stem cells
MX356756B (es) 2008-11-20 2018-06-11 Centocor Ortho Biotech Inc Células madre pluripotentes en microportadores.
EP2366022B1 (en) * 2008-11-20 2016-04-27 Janssen Biotech, Inc. Methods and compositions for cell attachment and cultivation on planar substrates
US20100209399A1 (en) * 2009-02-13 2010-08-19 Celavie Biosciences, Llc Brain-derived stem cells for repair of musculoskeletal system in vertebrate subjects
US8642334B2 (en) 2009-02-17 2014-02-04 Memorial Sloan Kettering Cancer Center Methods of neural conversion of human embryonic stem cells
US9109245B2 (en) 2009-04-22 2015-08-18 Viacyte, Inc. Cell compositions derived from dedifferentiated reprogrammed cells
DK2421957T3 (da) 2009-04-22 2021-01-25 Viacyte Inc Cellesammensætninger afledt af dedifferentierede omprogrammerede celler
US8785185B2 (en) 2009-07-20 2014-07-22 Janssen Biotech, Inc. Differentiation of human embryonic stem cells
JP5819825B2 (ja) 2009-07-20 2015-11-24 ヤンセン バイオテツク，インコーポレーテツド ヒト胚性幹細胞の分化
JP6219568B2 (ja) 2009-07-20 2017-10-25 ヤンセン バイオテツク，インコーポレーテツド ヒト胚性幹細胞の分化
WO2011017315A2 (en) 2009-08-03 2011-02-10 Recombinetics, Inc. Methods and compositions for targeted gene modification
EP2494035B1 (en) * 2009-10-29 2018-02-28 Janssen Biotech, Inc. Pluripotent stem cells
EP2501802A4 (en) 2009-11-17 2013-08-21 Advanced Cell Tech Inc METHOD FOR PRODUCING HUMAN RPE CELLS AND PHARMACEUTICAL PREPARATIONS FROM HUMAN RPE CELLS
MX343786B (es) * 2009-12-23 2016-11-22 Janssen Biotech Inc Diferenciacion de celulas madre embrionarias humanas.
RU2664864C1 (ru) * 2009-12-23 2018-08-23 Янссен Байотек, Инк. Способы увеличения экспрессии ngn3 и nkx6.1 в эндокринных клетках поджелудочной железы
JP6013196B2 (ja) * 2010-03-01 2016-10-25 ヤンセン バイオテツク，インコーポレーテツド 多能性幹細胞から誘導した細胞を精製するための方法
RU2587634C2 (ru) 2010-05-12 2016-06-20 Янссен Байотек, Инк. Дифференцирование эмбриональных стволовых клеток человека
WO2012009377A2 (en) 2010-07-12 2012-01-19 University Of Southern California Biocompatible substrate for facilitating interconnections between stem cells and target tissues and methods for implanting same
SG10201506264QA (en) 2010-08-12 2015-09-29 Janssen Biotech Inc Treatment of diabetes with pancreatic endocrine precursor cells
KR101851956B1 (ko) 2010-08-31 2018-04-25 얀센 바이오테크 인코포레이티드 인간 배아 줄기 세포의 분화
ES2659393T3 (es) 2010-08-31 2018-03-15 Janssen Biotech, Inc. Diferenciación de células madre embrionarias humanas
ES2585028T3 (es) 2010-08-31 2016-10-03 Janssen Biotech, Inc. Diferenciación de células madre pluripotentes
CA2814860C (en) 2010-10-22 2020-08-25 Biotime Inc. Methods of modifying transcriptional regulatory networks in stem cells
JP6002675B2 (ja) * 2010-12-17 2016-10-05 ビオラミナ アーベー 細胞培養培地
EP2801377B1 (en) 2011-03-04 2019-08-07 The Regents of The University of California Hydrogel comprising cells for local release of growth factors to mediate motor recovery after stroke
US8785190B2 (en) 2011-04-06 2014-07-22 Sanbio, Inc. Methods and compositions for modulating peripheral immune function
US8877489B2 (en) 2011-12-05 2014-11-04 California Institute Of Technology Ultrathin parylene-C semipermeable membranes for biomedical applications
US10478206B2 (en) 2011-04-29 2019-11-19 University Of Southern California Instruments and methods for the implantation of cell-seeded substrates
WO2012170853A1 (en) 2011-06-10 2012-12-13 Wisconsin Alumni Research Foundation ("Warf") Methods and devices for differentiating pluripotent stem cells into cells of the pancreatic lineage
WO2013010045A1 (en) 2011-07-12 2013-01-17 Biotime Inc. Novel methods and formulations for orthopedic cell therapy
CA2854578C (en) 2011-11-04 2023-01-03 Memorial Sloan-Kettering Cancer Center Midbrain dopamine (da) neurons for engraftment
US9248013B2 (en) 2011-12-05 2016-02-02 California Institute Of Technology 3-Dimensional parylene scaffold cage
RU2705001C2 (ru) 2011-12-22 2019-11-01 Янссен Байотек, Инк. Дифференцировка эмбриональных стволовых клеток человека в одногормональные инсулинположительные клетки
CA2866590A1 (en) 2012-03-07 2013-09-12 Janssen Biotech, Inc. Defined media for expansion and maintenance of pluripotent stem cells
KR20160027217A (ko) 2012-05-23 2016-03-09 에프. 호프만-라 로슈 아게 내배엽 및 간세포를 수득하고 사용하는 조성물 및 방법
JP6469003B2 (ja) 2012-06-08 2019-02-13 ヤンセン バイオテツク，インコーポレーテツド 膵内分泌細胞へのヒト胚性幹細胞の分化
CN102732477B (zh) * 2012-06-15 2013-06-19 江苏瑞思坦生物科技有限公司 人脂肪干细胞无血清基础培养基
KR102055438B1 (ko) 2012-07-31 2019-12-19 에이지엑스 쎄라퓨틱스, 인크. Hla g-변형된 세포 및 방법
AU2013237760A1 (en) 2012-10-08 2014-04-24 Biotime, Inc. Differentiated progeny of clonal progenitor cell lines
AU2013248265B2 (en) 2012-11-08 2018-11-01 Viacyte, Inc. Scalable primate pluripotent stem cell aggregate suspension culture and differentiation thereof
JP6557146B2 (ja) 2012-12-31 2019-08-07 ヤンセン バイオテツク，インコーポレーテツド 多能性幹細胞から膵臓内分泌細胞膵臓内分泌細胞への分化のための、空気−液体界面での、ヒト胚性幹細胞の培養
EP2938723B1 (en) 2012-12-31 2023-02-01 Janssen Biotech, Inc. Differentiation of human embryonic stem cells into pancreatic endocrine cells using hb9 regulators
US10370644B2 (en) 2012-12-31 2019-08-06 Janssen Biotech, Inc. Method for making human pluripotent suspension cultures and cells derived therefrom
EP4039798A1 (en) 2012-12-31 2022-08-10 Janssen Biotech, Inc. Suspension and clustering of human pluripotent cells
US8859286B2 (en) 2013-03-14 2014-10-14 Viacyte, Inc. In vitro differentiation of pluripotent stem cells to pancreatic endoderm cells (PEC) and endocrine cells
EP2989198A4 (en) 2013-04-26 2016-10-26 Sloan Kettering Inst Cancer CORTICAL INTERNEURONES AND OTHER NEURONAL CELLS PRODUCED BY DIRECTED DIFFERENTIATION OF PLURIPOTENT AND MULTIPOTENT CELLS
WO2014197421A1 (en) 2013-06-05 2014-12-11 Biotime, Inc. Compositions and methods for induced tissue regeneration in mammalian species
JP2016527878A (ja) 2013-06-14 2016-09-15 ザ ユニバーシティー オブ クイーンズランド 腎臓前駆細胞
EP3071685A4 (en) 2013-11-21 2017-07-05 Memorial Sloan Kettering Cancer Center Specification of functional cranial placode derivatives from human pluripotent stem cells
US11078462B2 (en) 2014-02-18 2021-08-03 ReCyte Therapeutics, Inc. Perivascular stromal cells from primate pluripotent stem cells
CA2949056A1 (en) 2014-05-16 2015-11-19 Janssen Biotech, Inc. Use of small molecules to enhance mafa expression in pancreatic endocrine cells
US10240127B2 (en) 2014-07-03 2019-03-26 ReCyte Therapeutics, Inc. Exosomes from clonal progenitor cells
CN104357379B (zh) * 2014-09-30 2017-08-08 刘兴宇 干细胞培养基
EP3387112A4 (en) 2015-12-07 2019-08-21 BioTime, Inc. METHODS FOR RETRANSTIGATION OF VARIOUS CELLS OF ADIPOSE BROWN TISSUE DERIVED FROM PLURIPOTENT STEM CELLS
EP3423158B1 (en) 2016-02-24 2023-11-15 The Rockefeller University Embryonic cell-based therapeutic candidate screening systems, models for huntington's disease and uses thereof
MA45479A (fr) 2016-04-14 2019-02-20 Janssen Biotech Inc Différenciation de cellules souches pluripotentes en cellules de l'endoderme de l'intestin moyen
KR101877793B1 (ko) * 2016-07-15 2018-07-13 주식회사 엔바이오텍 줄기세포 배양용 무혈청 배지 조성물 및 이를 이용한 줄기세포의 배양 방법
AU2017335841A1 (en) 2016-09-28 2019-04-04 Organovo, Inc. Use of engineered renal tissues in assays
CN106754652B (zh) * 2017-03-06 2019-04-02 广州润虹医药科技股份有限公司 iPS细胞分化成外胚层祖细胞的无血清诱导培养基及诱导方法
CN106754657B (zh) * 2017-03-28 2022-07-22 北京赛斯达生物技术有限公司 一种猴胚胎干细胞的无血清培养基
US20210123016A1 (en) 2018-05-02 2021-04-29 Novartis Ag Regulators of human pluripotent stem cells and uses thereof
CN114109294A (zh) * 2020-08-26 2022-03-01 中石化胜利石油工程有限公司管具技术服务中心 闸板拆装装置及其控制系统

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US670372A (en) * 1898-01-13 1901-03-19 William D Carpenter Process of producing casein products.
US5405772A (en) * 1993-06-18 1995-04-11 Amgen Inc. Medium for long-term proliferation and development of cells
US5453357A (en) * 1992-10-08 1995-09-26 Vanderbilt University Pluripotential embryonic stem cells and methods of making same
US5750376A (en) * 1991-07-08 1998-05-12 Neurospheres Holdings Ltd. In vitro growth and proliferation of genetically modified multipotent neural stem cells and their progeny

Family Cites Families (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5612211A (en) * 1990-06-08 1997-03-18 New York University Stimulation, production and culturing of hematopoietic progenitor cells by fibroblast growth factors
US5639046A (en) * 1992-07-21 1997-06-17 Fabio Perini S.P.A. Machine and method for the formation of coreless logs of web material
US5690926A (en) 1992-10-08 1997-11-25 Vanderbilt University Pluripotential embryonic cells and methods of making same
US5843780A (en) * 1995-01-20 1998-12-01 Wisconsin Alumni Research Foundation Primate embryonic stem cells
AU3392697A (en) 1996-06-14 1998-01-07 Regents Of The University Of California, The (in vitro) derivation and culture of primate pluripotent stem cells and therapeutic uses thereof
JP2001508302A (ja) 1997-01-10 2001-06-26 ライフ テクノロジーズ，インコーポレイテッド 胚性幹細胞血清置換
US6331406B1 (en) * 1997-03-31 2001-12-18 The John Hopkins University School Of Medicine Human enbryonic germ cell and methods of use
GB9722370D0 (en) * 1997-10-22 1997-12-17 Ici Plc Dye sheet cassette and printing apparatus
US6800480B1 (en) 1997-10-23 2004-10-05 Geron Corporation Methods and materials for the growth of primate-derived primordial stem cells in feeder-free culture
DE19756864C5 (de) 1997-12-19 2014-07-10 Oliver Brüstle Neurale Vorläuferzellen, Verfahren zu ihrer Herstellung und ihre Verwendung zur Therapie von neuralen Defekten
US6667176B1 (en) 2000-01-11 2003-12-23 Geron Corporation cDNA libraries reflecting gene expression during growth and differentiation of human pluripotent stem cells
US7410798B2 (en) * 2001-01-10 2008-08-12 Geron Corporation Culture system for rapid expansion of human embryonic stem cells
DE60026512D1 (de) 1999-05-07 2006-05-04 Univ Utah Res Found Auf eine zellinie beschränkte vorläuferzellen aus dem neuralrohr der maus und aus embryonalen stammzellen der maus
IL129966A (en) * 1999-05-14 2009-12-24 Technion Res & Dev Foundation ISOLATED HUMAN EMBRYOID BODIES (hEB) DERIVED FROM HUMAN EMBRYONIC STEM CELLS
US6750581B2 (en) * 2002-01-24 2004-06-15 Visteon Global Technologies, Inc. Automotive alternator stator assembly with rectangular continuous wire

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US670372A (en) * 1898-01-13 1901-03-19 William D Carpenter Process of producing casein products.
US5750376A (en) * 1991-07-08 1998-05-12 Neurospheres Holdings Ltd. In vitro growth and proliferation of genetically modified multipotent neural stem cells and their progeny
US5453357A (en) * 1992-10-08 1995-09-26 Vanderbilt University Pluripotential embryonic stem cells and methods of making same
US5405772A (en) * 1993-06-18 1995-04-11 Amgen Inc. Medium for long-term proliferation and development of cells

Also Published As

Publication number Publication date
IL151270A0 (en) 2003-04-10
CA2402299C (en) 2012-12-18
WO2001066697A3 (en) 2002-03-07
US7217569B2 (en) 2007-05-15
IS6515A (is) 2002-08-20
NO20024200L (no) 2002-09-03
MXPA02008698A (es) 2003-04-14
HK1053616A1 (en) 2003-10-31
IL151270A (en) 2008-07-08
NO335780B1 (no) 2015-02-16
AU4197301A (en) 2001-09-17
US20060040384A1 (en) 2006-02-23
CA2402299A1 (en) 2001-09-13
US7005252B1 (en) 2006-02-28
NO20024200D0 (no) 2002-09-03
JP2003525625A (ja) 2003-09-02
JP5839666B2 (ja) 2016-01-06
NZ520701A (en) 2004-03-26
EP1261691B1 (en) 2013-07-31
JP5717311B2 (ja) 2015-05-13
CN1416345A (zh) 2003-05-07
AU2001241973B2 (en) 2006-11-09
JP2011234735A (ja) 2011-11-24
BR0108507A (pt) 2002-12-17
EP1261691A2 (en) 2002-12-04
US20030190748A1 (en) 2003-10-09
JP2012005489A (ja) 2012-01-12
US20050148070A1 (en) 2005-07-07
WO2001066697A2 (en) 2001-09-13
KR100795760B1 (ko) 2008-01-21
KR20030032926A (ko) 2003-04-26

Similar Documents

Publication Publication Date Title
CN100372928C (zh) 灵长类动物胚胎干细胞的无血清培养物
Kanatsu-Shinohara et al. Improved serum-and feeder-free culture of mouse germline stem cells
KR101437927B1 (ko) 영장류 배아 줄기 세포의 배양
Rodin et al. Monolayer culturing and cloning of human pluripotent stem cells on laminin-521–based matrices under xeno-free and chemically defined conditions
Farini et al. Growth factors sustain primordial germ cell survival, proliferation and entering into meiosis in the absence of somatic cells
CA2307807C (en) Methods and materials for the growth of primate-derived primordial stem cells in feeder-free culture
US8211699B2 (en) Methods for culturing pluripotent stem cells in suspension using ERBB3 ligands
Amit et al. Feeder‐free culture of human embryonic stem cells
US20070010011A1 (en) Defined media for pluripotent stem cell culture
CN102395672A (zh) 用于干细胞培养的方法和组合物
CN102741396A (zh) 支持多能细胞生长的小分子和其方法
Ozawa et al. Culturing human pluripotent stem cells for regenerative medicine
CN101410510A (zh) 含激酶抑制剂的培养基及其应用
JP6446496B2 (ja) 霊長類胚性幹細胞の培養
AU2007200575B2 (en) Serum free cultivation of primate embryonic stem cells

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
REG Reference to a national code

Ref country code: HK

Ref legal event code: GR

Ref document number: 1053616

Country of ref document: HK

CX01 Expiry of patent term
CX01 Expiry of patent term

Granted publication date: 20080305