Kaur Jasmeet, Tilkins Mary Lynn, Eckert Richard, Boucher Shayne
Primary and Stem Cell Systems, Life Technologies, Carlsbad, CA, USA.
Methods Mol Biol. 2013;997:115-26. doi: 10.1007/978-1-62703-348-0_9.
Defined pluripotent stem cell culture media is a valuable tool for tracking and analyzing morphological, cell signaling and gene expression changes in human embryonic stem cells. Cultivation of hESC under xeno-free culture settings provides researchers with a consistent and reproducible environment to test experimental hypotheses and move towards translational and clinical research (Richards et al., Nat Biotechnol 20:933-936, 2002; Richards et al., Stem Cells 21:546-556, 2003). One of the primary concerns of the xenogeneic culture is the transfer of foreign pathogens or antigens that induce disease or immune response by the patient. These undesirable by-products may come from the use of murine-derived feeder cells, xenogeneic matrices, or from animal-derived components found in the cell culture medium or matrix used to isolate or expand the stem cells (Beattie et al., Stem Cells 23:489-495, 2005; Koivisto et al., Reprod Biomed Online 9:330-337, 2004). This chapter describes standardized protocols for obtaining consistent and reproducible results for culturing PSC under xeno-free, feeder-free, or feeder-based systems.
限定性多能干细胞培养基是追踪和分析人类胚胎干细胞形态、细胞信号传导及基因表达变化的宝贵工具。在无外源成分的培养条件下培养人胚胎干细胞，为研究人员提供了一个一致且可重复的环境，用于检验实验假设并推进转化研究和临床研究（Richards等人，《自然生物技术》20:933 - 936，2002年；Richards等人，《干细胞》21:546 - 556，2003年）。异种培养的一个主要问题是外来病原体或抗原的转移，这些物质会引发患者的疾病或免疫反应。这些不良副产物可能源于使用鼠源饲养细胞、异种基质，或存在于用于分离或扩增干细胞的细胞培养基或基质中的动物源性成分（Beattie等人，《干细胞》23:489 - 495，2005年；Koivisto等人，《生殖生物医学在线》9:330 - 337，2004年）。本章介绍了在无外源成分、无饲养层或有饲养层体系下培养多能干细胞以获得一致且可重复结果的标准化方案。
