[Isolation of viable karyo- and cyoplasts from the fertilized ova of mice using Ficoll-gradient centrifugation].
The fertilized mouse eggs obtained after the hormonal stimulation of the females were denuded by pronase and incubated for 1 h at 37 degrees in the culture medium with cytochalasin B (5 micrograms/ml) and, then, centrifuged 20 min (12,000 g) at ca. 31 degrees in Ficoll gradient added with cytochalasin B. Under these conditions the eggs were divided in anuclear (cytoplasts) and nuclear (karyoplasts) fragments. The cytoplasts kept their shape and ability for protein synthesis during cultivation and the karyoplasts performed one cell division. The viable cyto- and karyoplasts obtained from the eggs may be used in the experiments on cell hybridization and production of embryos-cybrids.
D000818:Animals; D002458:Cell Fractionation; D002459:Cell Fusion; D002467:Cell Nucleus / Q000648:ultrastructure*; D002469:Cell Separation; D002470:Cell Survival; D002499:Centrifugation, Density Gradient / Q000379:methods; D003571:Cytochalasin B / Q000494:pharmacology; D005260:Female; D005362:Ficoll; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D015053:Zygote / Q000166:cytology* / Q000187:drug effects
D005362:Ficoll; D003571:Cytochalasin B
Poluchenie zhiznesposobnykh kario- i tsitoplastov iz oplodotvorennykh iaĭtsekletok mysheĭ s pomoshch'iu tsentrifugirovaniia v gradiente fikolla.