Preferential loss of porcine chromosomes in reprogrammed interspecies cell hybrids.
Fusion of terminally differentiated somatic cells with pluripotent embryonic stem cells has been proposed as model for reprogramming the somatic cell genome, and may contribute to our understanding of the underlying mechanisms of this epigenetic process. We established an interspecies cell fusion model using murine embryonic stem cells (ESCs) and porcine fibroblasts. These inter-species fusion experiments yielded much lower conversion efficiency rates than murine intraspecies fusion. Nevertheless, two double-resistant mouse-pig hybrid clones could be generated. Reactivation of the porcine OCT4 gene, an essential pluripotent stem cell marker, and demethylation of the porcine OCT4 promoter in hybrid clone 1, suggested successful reprogramming of porcine chromosomes. A rapid loss of porcine chromosomes was observed during the selection phase. Spectral karyotyping (SKY) analysis showed that fusion-hybrid clone 1 carried a tetraploid mouse chromosome complement with only few pig chromosomes and/or chromosomal fragments. Hybrid clone 2 had a diploid set of murine chromosomes complements and also contained an interspecies chromosome fusion product. Interspecies cell fusion results in hybrid cells that retained the complement of mouse chromosomes and preferentially lose porcine chromosomes during colony expansion. Neither species-specific chromosomal segregation nor reprogrammed diploid porcine cells were observed. These findings indicate that generation of reprogrammed pluripotent diploid cells by cell fusion may require additional supporting provisions.
Institut of Farm Animal Genetics, Friedrich-Loeffler-Institute, Department of Biotechnology, Mariensee, Neustadt, Germany.
Monika
M
Nowak-Imialek
Wilfried A
WA
Kues
Cornelia
C
Rudolph
Brigitte
B
Schlegelberger
Ulrike
U
Taylor
Joseph W
JW
Carnwath
Heiner
H
Niemann
D000818:Animals; D030801:Animals, Genetically Modified; D002459:Cell Fusion / Q000379:methods; D002460:Cell Line; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D033481:Chromosomes, Mammalian; D004622:Embryo, Mammalian / Q000166:cytology; D053595:Embryonic Stem Cells / Q000502:physiology; D005240:Feasibility Studies; D006822:Hybrid Cells; D051379:Mice; D008822:Mice, Transgenic; D008954:Models, Biological; D039904:Pluripotent Stem Cells / Q000502:physiology; D034421:Sus scrofa / Q000235:genetics