Vitrification of intact and half 8-cell mouse embryos injected with DNA at 2-cell stage.
Two-cell embryos were collected from (C57BL/6 x CBA)F1 mice, microinjected with foreign DNA and transferred into temporary recipients. Intact and half 8-cell embryos recovered from oviducts 24 h later were frozen in microdrops of vitrification medium and stored in liquid nitrogen for 2 to 30 days. Development of frozen/thawed embryos was assessed by in vitro culture or transfer into day-1 recipients. From 1200 2-cell embryos 77% intact and 20% half embryos were obtained one hour after DNA injections. After transfer of 600 intact and 300 half embryos into temporary recipients, 77% intact and 73% half 8-cell embryos were recorded, respectively. Survival and in vitro development of frozen/thawed DNA-injected embryos were high, as 97% intact and 91% half 8-cell embryos developed to morulae or blastocysts. After transfer of 300 intact and 150 half 8-cell embryos into day-1 recipients, 99 (33%) and 19 (13%) young were born. None of the mice born from DNA-injected and cryopreserved embryos integrated injected genes.
Landa
V
V
Institute of Molecular Genetics, Czechoslovak Academy of Sciences, Praha.
Slezinger
M S
MS
D000818:Animals; D030801:Animals, Genetically Modified / Q000196:embryology; D015925:Cryopreservation / Q000379:methods; D004247:DNA / Q000235:genetics; D004622:Embryo, Mammalian; D005260:Female; D007267:Injections; D051379:Mice; D008815:Mice, Inbred Strains; D008846:Micromanipulation; D011247:Pregnancy; D014162:Transfection
D004247:DNA