GMP Scale-Up and Banking of Pluripotent Stem Cells for Cellular Therapy Applications
GMP Scale-Up and Banking of Pluripotent Stem Cells for Cellular Therapy Applications
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DOI：
10.1007/978-1-61779-201-4_11
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发表时间：
2011-01-01
期刊：
影响因子：
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通讯作者：
Couture, Larry A.
中科院分区：
文献类型：
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作者：
Ausubel, Lara J.;Lopez, Patricia M.;Couture, Larry A.
人类多能干细胞（PSCs）包括人类胚胎干细胞（ESCs）和诱导多能干细胞（iPSCs），是再生医学和人类早期发育研究中细胞治疗的重要来源。因此，开发大规模储存人类PSC系的方法变得越来越重要。有几种成熟的方法来繁殖人类psc。开发良好生产规范（GMP）库的关键是确定一种生产方法，该方法适用于使用有完整文件记录的材料进行大规模生产。我们已经利用动物饲养细胞、动物基质或无动物基质开发了几种hESCs库。在小鼠胚胎成纤维细胞（mef）上培养hESCs的方法已经建立，对产生研究级细胞库非常有帮助。由于研究实验室培养的大多数人类ESCs都暴露在异种试剂中，因此在制造主细胞库中使用的所有材料并不一定都是无动物来源的。然而，随着该领域的发展，毫无疑问，生产一种无异种试剂的临床使用细胞库将变得越来越重要，特别是可能含有对人类健康或细胞产品完整性有潜在风险的病毒的非人类饲养细胞。因此，即使对于先前暴露于异种试剂的细胞系，也必须尽量减少细胞系随后暴露于其他不确定试剂的情况。我们特别描述了在Matrigel（一种动物基质）和CELLstart（一种无动物基质）上培养hESCs的方法，这些方法可用于生产hESCs，作为临床制造过程的一部分。
Human pluripotent stem cells (PSCs), which include human embryonic stem cells (ESCs) as well as induced pluripotent stem cells (iPSCs), represent an important source of cellular therapies in regenerative medicine and the study of early human development. As such, it is becoming increasingly important to develop methods for the large-scale banking of human PSC lines. There are several well-established methods for the propagation of human PSCs. The key to development of a good manufacturing practice (GMP) bank is to determine a manufacturing method that is amenable to large-scale production using materials that are fully documented. We have developed several banks of hESCs using animal feeder cells, animal-based matrices, or animal-free matrices. Protocols for growing hESCs on mouse embryonic fibroblasts (MEFs) are well established and are very helpful for producing research grade banks of cells. As most human ESCs cultured by research laboratories have been exposed to xenogeneic reagents, it is not imperative that all materials used in the production of a master cell bank be animal-free in origin. Nevertheless, as the field develops, it will no doubt become increasingly important to produce a bank of cells for clinical use without xenogeneic reagents, particularly nonhuman feeder cells which might harbor viruses with potential risk to human health or cell product integrity. Thus, even for cell lines previously exposed to xenogeneic reagents, it is important to minimize any subsequent exposure of the cell lines to additional adventitious agents. We have specifically described procedures for the growth of hESCs on Matrigel, an animal-matrix, and CELLstart, an animal-free matrix, and these can be used to produce hESCs as part of a clinical manufacturing process.