The Comparison between Conditioned Media and Serum-Free Media in Human Embryonic Stem Cell Culture and Differentiation
The Comparison between Conditioned Media and Serum-Free Media in Human Embryonic Stem Cell Culture and Differentiation
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DOI：
10.1089/cell.2009.0099
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发表时间：
2010-04-01
影响因子：
1.6
通讯作者：
Hay, David C.
中科院分区：
文献类型：
--
作者：
Hannoun, Zara;Fletcher, Judy;Hay, David C.
人类胚胎干细胞（hESCs）通过其自我更新的能力，同时保持多能性，为人类体细胞类型提供了取之不尽的供应。因此，hesc衍生的细胞类型对于从体外建模到治疗用途的应用非常重要。然而，为了充分发挥其潜力，未分化细胞及其衍生物的生长必须在规定的培养条件下进行。许多研究小组使用小鼠胚胎成纤维细胞（MEF）和MEF条件培养基（CM）维持hESCs。在发展hESC技术中，使用小鼠系统来支持hESC是必不可少的；然而，它们受到一些主要限制，包括缺乏定义、异种性质、批次之间的差异和劳动密集型生产。因此，如果要保证hESC系及其衍生物的质量并按照GMP生产，hESC培养定义是必不可少的。我们已经启动了标准化hESC组织培养的过程，并采用了两种无血清培养基：mTeSR （MT）和Stem Pro （SP）。使用MT和SP将hESCs维持在多能状态超过30代。此外，我们提供的证据表明，在MT和SP中维持的hESCs产生了与CM观察到的相同水平的人肝内胚层。这些数据表明，MT和SP是hESC培养中MEF-CM的有效替代品，有助于hESC体外模型的标准化和最终的应用。
Human embryonic stem cells (hESCs) offer an inexhaustible supply of human somatic cell types through their ability to self-renew while retaining pluripotency. As such, hESC-derived cell types are important for applications ranging from in vitro modeling to therapeutic use. However, for their full potential to be realized, both the growth of the undifferentiated cells and their derivatives must be performed in defined culture conditions. Many research groups maintain hESCs using mouse embryonic fibroblasts (MEF) and MEF conditioned medium (CM). The use of murine systems to support hESCs has been imperative in developing hESC technology; however, they suffer from some major limitations including lack of definition, xenobiotic nature, batch-to-batch variation, and labor-intensive production. Therefore, hESC culture definition is essential if hESC lines, and their derivatives are to be quality assured and manufactured to GMP. We have initiated the process of standardizing hESC tissue culture and have employed two serum-free media: mTeSR (MT) and Stem Pro (SP). hESCs were maintained in a pluripotent state, for over 30 passages using MT and SP. Additionally, we present evidence that hESCs maintained in MT and SP generate equivalent levels of human hepatic endoderm as observed with CM. This data suggests that MT and SP are effective replacements for MEF-CM in hESC culture, contributing to the standardization of hESC in vitro models and ultimately their application.