Genome-wide analysis of gene expression during somatic cell reprogramming

Reprogrammed mouse embryonic fibroblasts generated clonal cell lines were analysed during maintained expression of the four reprogramming transcription factors (C-Myc, Oct4, Sox2, Klf4). The transgene expressing clonal cell lines segregated in to two Nanog positive cell types, F-class and C-class, that both differentiate in to three germ layers. The emergence of F-class cells is dependent on the expression of all four factors with a necessity for high levels of transgene expression. The F-class cells represent a novel and stable cell state. Mouse embryonic fibroblasts were reprogrammed using four transcription factors (C-Myc, Oct4, Sox2, Klf4) and clonal cell lines established. Cell lines were maintained in doxycycline supplemented media to sustain transgene expression. Global gene expression profile analysis was performed on the clonal cell lines at day16, with respect to embryonic stem cells and parental fibroblast cells. F-class cells (clone 1) were maintained for a further two weeks and found not to acquire expression of ESC associated genes.