The Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, Queensland 4072, Australia.
Biomaterials. 2010 Nov;31(32):8281-8. doi: 10.1016/j.biomaterials.2010.07.037. Epub 2010 Aug 2.
Human embryonic stem cells (hESC) are expected to provide revolutionary therapeutic applications and drug discovery technologies. In order for this to be achieved a reproducible, defined animal component free culture system is required for the scale-up production of undifferentiated hESC. In this work we have investigated the applicability of a recombinantly produced domain of human vitronectin as an extracellular matrix alternative to the common standards Geltrex or Matrigel. In addition we have validated an ascorbate free media capable of supporting CD30(low) populations of hESC through a multi-factorial analysis of bFGF and Activin A. The recombinant vitronectin domain combined with the ascorbate free media were capable of supporting 3 cell lines, MEL1, MEL2 and hES3 for 10 or more passages while maintaining hESC pluripotency markers and differentiation capacity. The culture method outlined here provides a platform for future investigation into growth factor and extracellular matrix effects on hESC maintenance prior to bioreactor scale-up.
人胚胎干细胞（hESC）有望提供革命性的治疗应用和药物发现技术。为了实现这一目标，需要一种可重复的、无定义的动物成分的培养系统，以便大规模生产未分化的 hESC。在这项工作中，我们研究了重组人 vitronectin 结构域作为细胞外基质替代物的适用性，以替代常见的 Geltrex 或 Matrigel 标准。此外，我们通过对 bFGF 和 Activin A 的多因素分析，验证了一种无抗坏血酸的培养基，该培养基能够支持 CD30(low) 群体的 hESC。重组 vitronectin 结构域与无抗坏血酸的培养基相结合，能够支持 MEL1、MEL2 和 hES3 细胞系传代 10 次以上，同时保持 hESC 多能性标记和分化能力。这里概述的培养方法为在生物反应器放大之前研究生长因子和细胞外基质对 hESC 维持的影响提供了一个平台。
