Nuclear reprogramming of somatic cells after fusion with human embryonic stem cells.
We have explored the use of embryonic stem cells as an alternative to oocytes for reprogramming human somatic nuclei. Human embryonic stem (hES) cells were fused with human fibroblasts, resulting in hybrid cells that maintain a stable tetraploid DNA content and have morphology, growth rate, and antigen expression patterns characteristic of hES cells. Differentiation of hybrid cells in vitro and in vivo yielded cell types from each embryonic germ layer. Analysis of genome-wide transcriptional activity, reporter gene activation, allele-specific gene expression, and DNA methylation showed that the somatic genome was reprogrammed to an embryonic state. These results establish that hES cells can reprogram the transcriptional state of somatic nuclei and provide a system for investigating the underlying mechanisms.
Cowan
Chad A
CA
Howard Hughes Medical Institute, Harvard Stem Cell Institute, Department of Molecular and Cellular Biology, Harvard University, 7 Divinity Avenue, Cambridge, MA 02138, USA.
Atienza
Jocelyn
J
Melton
Douglas A
DA
Eggan
Kevin
K
D000328:Adult; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002453:Cell Cycle; D002454:Cell Differentiation; D002459:Cell Fusion; D002460:Cell Line; D002467:Cell Nucleus / Q000502:physiology*; D048430:Cell Shape; D017690:Cell Transplantation; D002877:Chromosomes, Human / Q000235:genetics; D004622:Embryo, Mammalian / Q000166:cytology*; D044127:Epigenesis, Genetic; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology*; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D006822:Hybrid Cells / Q000166:cytology / Q000502:physiology*; D008297:Male; D051379:Mice; D008819:Mice, Nude; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000502:physiology*; D011123:Polyploidy; D013724:Teratoma / Q000473:pathology; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014162:Transfection
D015415:Biomarkers