Bergström Rosita, Ström Susanne, Holm Frida, Feki Anis, Hovatta Outi
Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden.
Methods Mol Biol. 2011;767:125-36. doi: 10.1007/978-1-61779-201-4_9.
Stem cell culture systems that rely on undefined animal-derived components introduce variability to the cultures and complicate their therapeutic use. The derivation of human embryonic stem cells and the development of methods to produce induced pluripotent stem cells combined with their potential to treat human diseases have accelerated the drive to develop xenogenic-free, chemically defined culture systems that support pluripotent self-renewal and directed differentiation. In this chapter, we describe four xeno-free culture systems that have been successful in supporting undifferentiated growth of hPSCs as well as methods for xeno-free subculture and cryopreservation of hPSCs. Each culture system consists of a xeno-free growth medium and xeno-free substratum: (1) TeSR2™ with human recombinant laminin (LN-511); (2) NutriStem™ with LN-511; (3) RegES™ with human foreskin fibroblasts (hFFs); (4) KO-SR Xeno-Free™/GF cocktail with CELLstart™ matrix.
依赖于成分不明确的动物源成分的干细胞培养系统会给培养带来变异性，并使其治疗用途变得复杂。人类胚胎干细胞的衍生以及诱导多能干细胞生产方法的发展，再加上它们治疗人类疾病的潜力，加速了开发无外源、化学成分明确的培养系统的进程，这些系统支持多能自我更新和定向分化。在本章中，我们描述了四种在支持人多能干细胞未分化生长方面取得成功的无外源培养系统，以及人多能干细胞的无外源传代培养和冷冻保存方法。每种培养系统都由无外源生长培养基和无外源基质组成：(1)含有重组人层粘连蛋白（LN-511）的TeSR2™；(2)含有LN-511的NutriStem™；(3)含有人类包皮成纤维细胞（hFFs）的RegES™；(4)含有CELLstart™基质的KO-SR无外源™/GF鸡尾酒。
