Yao Shuyuan, Chen Shuibing, Clark Julie, Hao Ergeng, Beattie Gillian M, Hayek Alberto, Ding Sheng
Department of Chemistry, The Scripps Research Institute, SR202, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Proc Natl Acad Sci U S A. 2006 May 2;103(18):6907-12. doi: 10.1073/pnas.0602280103. Epub 2006 Apr 21.
Chemically defined medium (CDM) conditions for controlling human embryonic stem cell (hESC) fate will not only facilitate the practical application of hESCs in research and therapy but also provide an excellent system for studying the molecular mechanisms underlying self-renewal and differentiation, without the multiple unknown and variable factors associated with feeder cells and serum. Here we report a simple CDM that supports efficient self-renewal of hESCs grown on a Matrigel-coated surface over multiple passages. Expanded hESCs under such conditions maintain expression of multiple hESC-specific markers, retain the characteristic hESC morphology, possess a normal karyotype in vitro, as well as develop teratomas in vivo. Additionally, several growth factors were found to selectively induce monolayer differentiation of hESC cultures toward neural, definitive endoderm/pancreatic and early cardiac muscle cells, respectively, in our CDM conditions. Therefore, this CDM condition provides a basic platform for further characterization of hESC self-renewal and directed differentiation, as well as the development of novel therapies.
用于控制人类胚胎干细胞（hESC）命运的化学成分确定的培养基（CDM）条件，不仅将促进hESC在研究和治疗中的实际应用，还将提供一个出色的系统来研究自我更新和分化背后的分子机制，而没有与饲养层细胞和血清相关的多种未知和可变因素。在此，我们报告一种简单的CDM，它支持在基质胶包被的表面上生长的hESC在多次传代过程中高效自我更新。在这种条件下扩增的hESC维持多种hESC特异性标志物的表达，保留hESC的特征形态，在体外具有正常的核型，并且在体内形成畸胎瘤。此外，在我们的CDM条件下，发现几种生长因子分别选择性地诱导hESC培养物向神经、定形内胚层/胰腺和早期心肌细胞的单层分化。因此，这种CDM条件为进一步表征hESC自我更新和定向分化以及开发新疗法提供了一个基础平台。
