Cell fusion is one approach that has been used to demonstrate nuclear reprogramming of somatic cells to a pluripotent-like state and is a useful tool for screening factors involved in reprogramming. Recent cell fusion studies reported that the overexpression of Nanog and SalI could improve the efficiency of reprogramming, whereas AID was shown to be essential for DNA demethylation and initiation of reprogramming. The aim of this study was to investigate factors affecting the reprogramming efficiency following cell fusion.
View Article and Find Full Text PDFEctopic expression of key reprogramming transgenes in somatic cells enables them to adopt the characteristics of pluripotency. Such cells have been termed induced pluripotent stem (iPS) cells and have revolutionized the field of somatic cell reprogramming, as the need for embryonic material is obviated. One of the issues facing both the clinical translation of iPS cell technology and the efficient derivation of iPS cell lines in the research laboratory is choosing the most appropriate somatic cell type for induction.
View Article and Find Full Text PDFThe ectopic expression of the key transcription factors Oct4, Sox2, c-Myc, and Klf-4 have been shown to reprogram somatic cells to a pluripotent state. In turn these induced pluripotent stem (iPS) cells, like embryonic stem (ES) cells, have been shown to be able to reprogram somatic cells by cell fusion. In this study we compare the differences and similarities between ES and iPS cells measured by somatic cell fusion to somatic cells harboring an Oct4-GFP transgene.
View Article and Find Full Text PDFIn this study we examine whether a somatic cell, once returned to a pluripotent state, gains the ability to reprogram other somatic cells. We reprogrammed mouse embryonic fibroblasts by viral induction of oct4, sox2, c-myc, and klf-4 genes. Upon fusion of the resulting iPS cells with somatic cells harboring an Oct4-GFP transgene we observed, GFP expression along with activation of Oct4 from the somatic genome, expression of key pluripotency genes, and positive immunostaining for Oct4, SSEA-1, and alkaline phosphatase.
View Article and Find Full Text PDFEven though the technique of mammalian SCNT is just over a decade old it has already resulted in numerous significant advances. Despite the recent advances in the reprogramming field, SCNT remains the bench-mark for the generation of both genetically unmodified autologous pluripotent stem cells for transplantation and for the production of cloned animals. In this review we will discuss the pros and cons of SCNT, drawing comparisons with other reprogramming methods.
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