Transcriptome analysis of immediate early events in mRNA-mediated cellular reprogramming of human foreskin fibroblasts. Homo sapiens

In this study human neonatal foreskin fibroblasts (HFF1 and BJ) were lipofected with an equal mixture of human reprogramming factor-encoding mRNAs. The cells were lysed and total RNA was harvested 24 hours post-transfection. Gene regulation was evaluated with respect to corresponding mock-transfected, Lipofectamine-treated control fibroblasts. For comparison, wildtype (un-treated) HFF1 and BJ fibroblasts as well as undifferentiated HFF1- and BJ-derived induced pluripotent stem cells and human embryonic stem cells (generated and maintained in our laboratory (refer to GSE26575) were included in the analysis. Overall design: Total RNA obtained from untreated human neonatal foreskin fibroblasts, reprogramming factor- and mock(Lipofectamine-treated)-transfected human neonatal fibroblasts, undifferentiated hESCs and iPSCs (derived from human neonatal foreskin fibroblasts).

本研究中，将人类新生儿包皮成纤维细胞（HFF1与BJ株）以等比例混合的编码人类重编程因子的信使核糖核酸（mRNA）进行脂质转染。转染24小时后裂解细胞并收集总RNA。以相应的仅经Lipofectamine处理的空白转染成纤维细胞为对照，评估基因调控情况。

为便于对照分析，本研究同时纳入未经处理的野生型HFF1及BJ成纤维细胞、未分化的HFF1与BJ来源诱导多能干细胞（induced pluripotent stem cells，iPSCs），以及本实验室构建并维持培养的人类胚胎干细胞（human embryonic stem cells，hESCs，相关信息参见GSE26575），将其纳入本次分析。

整体实验设计：本分析所用总RNA来源于以下样本：未经处理的人类新生儿包皮成纤维细胞、经重编程因子转染的人类新生儿包皮成纤维细胞、经空白（Lipofectamine处理）转染的人类新生儿包皮成纤维细胞，以及由人类新生儿包皮成纤维细胞诱导得到的未分化人类胚胎干细胞与诱导多能干细胞。