Functional full-term placentas formed from parthenogenetic embryos using serial nuclear transfer.
Mammalian parthenogenetic embryos invariably die in mid-gestation from imprinted gene defects and placental hypoplasia. Based on chimera experiments, trophoblastic proliferation is supposed to be inhibited in the absence of a male genome. Here, we show that parthenogenetic mouse embryonic cell nuclei can be reprogrammed by serial rounds of nuclear transfer without using any genetic modification. The durations of survival in uteri of cloned foetuses derived from green fluorescent protein (GFP)-labelled parthenogenetic cell nuclei were extended with repeated nuclear transfers. After five repeats, live cloned foetuses were obtained up to day 14.5 of gestation; however, they did not survive longer even when we repeated nuclear transfer up to nine times. All foetuses showed intestinal herniation and possessed well-expanded large placentas. When embryonic stem (ES) cells derived from fertilised embryos were aggregated with the cloned embryos, full-term offspring with large placentas were obtained from the chimeric embryos. Those placentas were derived from parthenogenetic cell nuclei, judging from GFP expression. The patterns of imprinted gene expression and methylation status were similar to their parthenogenetic origin, except for Peg10, which showed the same level as in the normal placenta. These results suggest that there is a limitation for foetal development in the ability to reprogramme imprinted genes by repeated rounds of nuclear transfer. However, the placentas of parthenogenetic embryos can escape epigenetic regulation when developed using nuclear transfer techniques and can support foetal development to full gestation.
Hikichi
Takafusa
T
RIKEN Center for Developmental Biology, Minatojima-minamimachi Chuo-ku, Kobe, Japan.
Ohta
Hiroshi
H
Wakayama
Sayaka
S
Wakayama
Teruhiko
T
D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation; D002460:Cell Line; D065150:Cellular Reprogramming / Q000235:genetics / Q000502:physiology; D019976:Cloning, Organism; D019175:DNA Methylation; D017931:DNA Primers / Q000235:genetics; D047108:Embryonic Development; D053595:Embryonic Stem Cells / Q000166:cytology; D005260:Female; D015870:Gene Expression; D018392:Genomic Imprinting; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008813:Mice, Inbred ICR; D008822:Mice, Transgenic; D053652:Nuclear Transfer Techniques; D010312:Parthenogenesis / Q000235:genetics / Q000502:physiology*; D010920:Placenta / Q000166:cytology / Q000502:physiology; D010929:Placentation; D039904:Pluripotent Stem Cells / Q000166:cytology; D011247:Pregnancy; D039901:Totipotent Stem Cells / Q000166:cytology; D018183:Transplantation Chimera
D017931:DNA Primers