Stem Cell Laboratory, School of Psychiatry, University of New South Wales, Sydney, Australia.
Stem Cells Dev. 2012 Jan 20;21(2):206-16. doi: 10.1089/scd.2010.0440. Epub 2011 Jun 1.
Use of animal feeder layers and serum containing media in the derivation and propagation of induced pluripotent stem cells (iPSCs) can hinder clinical translation, because of the presence of xeno-material/pathogens. A defined and standardized system would be ideal for generating a homogenous population of iPSCs, which closely resembles human embryonic stem cells (hESCs). This article presents a novel and extensive comparison between in-house produced iPSCs and hESCs under "feeder" and "feeder-free" conditions, using transcriptomic genome-wide microarray analysis. We generated a list of pluripotency-associated and bivalent domain-containing genes by meta-analysis to measure qualitatively the degree of reprogramming in feeder-free derived iPSCs, in which both profiles displayed similar levels of gene expression as in hESCs. Gene ontology analysis showed that feeder-free iPSCs have enriched terms belonging to DNA repair/replication and cell cycle, which are signature to pluripotent cells. Transcriptomic data combined with directed differentiation assays, indicated that variability among iPSC lines is minimized when using a feeder-free cultural system, which may serve as a platform for further developing regenerative medicine compliant human iPSCs.
在诱导多能干细胞（iPSCs）的衍生和繁殖中使用含有动物饲养层和血清的培养基会阻碍临床转化，因为其中存在异源材料/病原体。一个定义明确且标准化的系统对于产生类似于人类胚胎干细胞（hESCs）的同质 iPSC 群体是理想的。本文通过转录组全基因组微阵列分析，在“饲养层”和“无饲养层”条件下对内部生产的 iPSC 和 hESC 进行了广泛的比较。我们通过荟萃分析生成了一组与多能性相关和包含二价结构域的基因列表，以定性地衡量无饲养层衍生的 iPSC 中的重编程程度，其中这两种谱都显示出与 hESC 相似的基因表达水平。基因本体论分析表明，无饲养层 iPSC 具有丰富的 DNA 修复/复制和细胞周期相关术语，这是多能细胞的特征。转录组数据结合定向分化实验表明，当使用无饲养层培养系统时，iPSC 系之间的变异性最小，这可能成为进一步开发符合再生医学要求的人类 iPSC 的平台。
