Reprogramming of somatic cells after fusion with induced pluripotent stem cells and nuclear transfer embryonic stem cells.
In 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. The iPS-somatic hybrids had the ability to differentiate into cell types indicative of the three germ layers and were able to localize to the inner cell mass of aggregated embryos. Furthermore, ntES cells were used as fusion partners to generate hybrids, which were also confirmed to be reprogrammed to a pluripotent state. These results demonstrate that once a somatic cell nucleus is reprogrammed, it acquires the capacity and potency to reprogram other somatic cells by cell fusion and shares this functional property with normal embryonic stem (ES) cells.
Sumer
Huseyin
H
Monash Institute of Medical Research, Monash University, Clayton, Victoria, Australia. huseyin.sumer@med.monash.edu.au
Jones
Karen L
KL
Liu
Jun
J
Heffernan
Corey
C
Tat
Pollyanna A
PA
Upton
Kyle R
KR
Verma
Paul J
PJ
D000818:Animals; D002459:Cell Fusion; D017690:Cell Transplantation / Q000379:methods; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D004622:Embryo, Mammalian / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006822:Hybrid Cells / Q000166:cytology* / Q000378:metabolism; D007150:Immunohistochemistry; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D008297:Male; D051379:Mice; D016513:Mice, SCID; D008822:Mice, Transgenic; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D011247:Pregnancy; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013724:Teratoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D014161:Transduction, Genetic
D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D049452:Green Fluorescent Proteins