优化生理性无动物源分子定义培养基和基质以维持人胚胎干细胞多能性。

Optimization of physiological xenofree molecularly defined media and matrices to maintain human embryonic stem cell pluripotency.

作者信息

Peiffer Isabelle, Barbet Romain, Hatzfeld Antoinette, Li Ma-Lin, Hatzfeld Jacques A

机构信息

Centre National de la Recherche Scientifique, Institut André Lwoff, Villejuif, France.

出版信息

Methods Mol Biol. 2010;584:97-108. doi: 10.1007/978-1-60761-369-5_5.

DOI:10.1007/978-1-60761-369-5_5

PMID:19907973

Abstract

We describe in this chapter the development of a xenofree molecularly defined medium, SBX, associated with xenofree matrices, to maintain human embryonic stem cell (hESC) pluripotency as determined by phenotypic, functional and TLDA studies. This simple, inexpensive, and more physiological culture condition has been chosen because (1) it is xenofree and molecularly defined; it is devoid of albumin, which is a carrier of undefined molecules; (2) it maintains pluripotency, but very significantly reduces differentiation gene expression during hESC self-renewal, as compared to the widely used culture conditions tested so far; and (3) it can be further improved by replacing high concentrations of expensive additives by physiological concentrations of new factors. Xenofree molecularly defined media and matrices represent valuable tools for elucidating still unknown functions of numerous embryonic genes using more physiological culture conditions. These genes encode potential new factors controlling hESC self-renewal and pluripotency.

摘要

在本章中，我们描述了一种无动物源的、分子成分明确的培养基SBX的研发情况，该培养基与无动物源基质相关联，用于维持人胚胎干细胞（hESC）的多能性，这是通过表型、功能和TLDA研究确定的。选择这种简单、廉价且更符合生理状态的培养条件是因为：（1）它是无动物源且分子成分明确的；它不含白蛋白，白蛋白是未定义分子的载体；（2）它能维持多能性，但与目前测试的广泛使用的培养条件相比，在hESC自我更新过程中能非常显著地降低分化基因的表达；（3）通过用新因子的生理浓度替代高浓度的昂贵添加剂，它还可以进一步改进。无动物源的、分子成分明确的培养基和基质是利用更符合生理状态的培养条件阐明众多胚胎基因仍未知功能的宝贵工具。这些基因编码控制hESC自我更新和多能性的潜在新因子。