Production of mice derived entirely from embryonic stem cells after injecting the cells into heat treated blastocysts.
The sensitivity of the inner cell mass (ICM) and trophectoderm (TE) of mouse blastocysts to high temperatures was examined. When blastocysts with a diameter of 100 to 120 microm treated for 15 to 20 min at 45 degrees C were cultured in vitro, the cell number in the ICM did not increase, although that in the TE did increase. After transfer of treated blastocysts to recipients, implantation was not drastically inhibited but no live fetuses were obtained. These results demonstrated that the ICM at the blastocyst stage was more sensitive to high temperature than the TE. ICM clumps or ES cells were injected into blastocysts treated for 20 min at 45 degrees C. After transfer of injected blastocysts to recipients, we obtained mice derived completely from ICM or ES cells as judged by GPI analysis. Since 4 of 7 ES-cell derived mice, but none of the 6 mice derived from the ICM died after birth, an as yet unidentified epigenetic alteration might have occurred during the establishment and/or culture of ES cells.
Amano
T
T
Laboratory of Animal Reproduction, College of Agriculture, Kinki University, Nara, Japan.
Nakamura
K
K
Tani
T
T
Kato
Y
Y
Tsunoda
Y
Y
D000818:Animals; D001755:Blastocyst; D005260:Female; D005298:Fertility; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary*; D006358:Hot Temperature; D008297:Male; D051379:Mice; D012621:Seasons; D013234:Stem Cells