title: The efficiency of cell fusion-based reprogramming is affected by the somatic cell type and the in vitro age of somatic cells.
abstract: 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. We conducted fusions of mouse embryonic stem cells (ESCs) with somatic cells carrying a GFP transgene under control of the Oct4 promoter (Oct4-GFP), which is normally repressed in nonpluripotent cells. The effect of somatic cell type on the reprogramming efficiency was investigated using Oct4-GFP expression as an indicator. Different somatic cell types were tested including mesenchymal stem cells (MSCs), adipose tissue-derived cells (ADCs), neural stem cells (NSCs), and these were compared with the mouse embryonic fibroblast (mEF) standard. The reprogramming efficiencies differed greatly, with mEFs (0.477 ± 0.003%) and MSCs (0.313 ± 0.003%) showing highest efficiencies while NSCs (0.023 ± 0.014%), and ADCs (0.006 ± 0.006%) had significantly lower reprogramming efficiencies (p < 0.05). The differences in the reprogramming efficiencies observed could be in part explained by the in vitro age of the somatic cells used. We demonstrated that the reprogramming efficiency of early passage mEFs was significantly higher compared with late passage mEFs (0.330 ± 0.166% vs. 0.021 ± 0.011%, p < 0.05), suggesting that senescence can affect reprogramming potential. In summary, this study shows that different somatic cell types do not have equivalent potential to be reprogrammed following fusion with ESCs. Furthermore, the in vitro age of somatic cells can also affect the reprogrammability of somatic cells. These findings constitute an important consideration for reprogramming studies.
authors: Centre for Reproduction and Development, Monash Institute of Medical Research, Monash University, Australia.
authors: Pollyanna Agnes
authors: PA
authors: Tat
authors: Huseyin
authors: H
authors: Sumer
authors: Daniele
authors: D
authors: Pralong
authors: Paul John
authors: PJ
authors: Verma
mesh_terms: D000818:Animals; D002459:Cell Fusion; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology*; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D019076:Transgenes
chemical_list: D050814:Octamer Transcription Factor-3; D011993:Recombinant Fusion Proteins