Multipotentiality of neuronal cells after spontaneous fusion with embryonic stem cells and nuclear reprogramming in vitro.
Primary mouse brain cells were cultured with HPRT (hypoxanthine phosphoribosyl transferase)-deficient ES (embryonic stem) cells to see if the ES cells could provide cues sufficient to reprogram a pluripotential state. After 5 days of coculture, HPRT-deficient ES cells were killed by selection in HAT (hypoxanthine, aminopterin, thymidine) medium. We observed islands of HAT-resistant ES-like cells surrounded by differentiated cells. Cell lines generated from three such "islands" proved to be spontaneous, pluripotential ES-neural hybrids, and gave rise to a chimera following blastocyst injection. Re-expression of the ES-specific gene Foxd3 from somatic-derived chromosomes suggested that the somatic nucleus had been reprogrammed. Our results raise the intriguing possibility that ASCs shown to contribute to multiple tissues in blastocyst-injection studies may not contribute as a result of pluripotency. Instead contributions may arise from spontaneous fusion events in which phenotype is determined by either cytoplasmic dominance, nuclear reprogramming, or both.
Pells
Steve
S
Roslin Institute (Edinburgh), Roslin, Midlothian, United Kingdom.
Di Domenico
Alexandra I
AI
Gallagher
Ed J
EJ
McWhir
Jim
J
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002459:Cell Fusion; D002460:Cell Line; D019070:Cell Lineage / Q000502:physiology; D002678:Chimera; D018920:Coculture Techniques; D006822:Hybrid Cells / Q000166:cytology / Q000502:physiology*; D007621:Karyotyping; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*