Effective production of microinjectable blastocysts for germ-line transmission of embryonic stem cells.
Blastocysts of C57BL/6 mice obtained either 2.5 or 3.5 days post-coitum (dpc) were examined for efficient microinjection after overnight in vitro culture. Incidences of zona-free embryos were much higher at 3.5 dpc after natural mating (1.05/mouse) and superovulation (2.83/mouse) than at 2.5 dpc after natural mating (0.05/mouse) and superovulation (0.01/mouse). By testing germ-line competency of gene-targeted J1 embryonic stem cells, superovulation and/or in vitro culture should be recommended for producing microinjectable blastocysts for production of high germ-line chimeras.
Kim
Sun-Uk
SU
Laboratory of Human Genomics, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Korea.
Han
Ying-Hao
YH
Lee
Tae-Hoon
TH
Hyun
Byung-Hwa
BH
Lee
Sang-Ho
SH
Lee
Dong-Seok
DS
Yu
Dae-Yeul
DY
D000818:Animals; D001755:Blastocyst / Q000166:cytology; D002478:Cells, Cultured; D002678:Chimera; D004624:Embryo Transfer; D005260:Female; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008845:Microinjections; D011247:Pregnancy; D013234:Stem Cells; D013480:Superovulation