Vallier Ludovic, Pedersen Roger
Department of Surgery and Cambridge Institute for Medical Research, Addenbrooke's Hospital, University of Cambridge, Cambridge, United Kingdom.
Curr Protoc Stem Cell Biol. 2008 Mar;Chapter 1:Unit 1D.4.1-1D.4.7. doi: 10.1002/9780470151808.sc01d04s4.
Generating fully functional differentiated cells from human embryonic stem cells and achieving this goal using clinically compatible conditions remain major challenges for the stem cell field. The presence of undefined components in standard culture media and protocols (including animal-derived serum, feeder cells, and extracellular matrices) has significantly impeded the achievement of these objectives. Here, we describe culture conditions to differentiate pluripotent cells in adherent conditions and in the absence of stroma cells, feeder cells, conditioned medium, serum, or complex matrices. Importantly, these defined culture conditions are devoid of animal products, thereby eliminating factors that could obscure analysis of developmental mechanisms or render the resulting tissues incompatible with future clinical applications.
从人类胚胎干细胞中生成功能完全分化的细胞,并在临床兼容的条件下实现这一目标,仍然是干细胞领域的主要挑战。标准培养基和方案中存在不确定成分(包括动物来源的血清、饲养细胞和细胞外基质),这严重阻碍了这些目标的实现。在这里,我们描述了在无饲养层细胞、条件培养基、血清或复杂基质的贴壁条件下分化多能细胞的培养条件。重要的是,这些明确的培养条件不含动物产品,从而消除了可能模糊发育机制分析或使所得组织与未来临床应用不兼容的因素。