[Modification of a method of cell enucleation using cytochalasin B and the study of cytoplast viability].
The modification of Prescott's (Prescott et al., 1972) method of enucleation in vitro was described. A special teflon chamber faciliatating the enucleation of monolayer cultured cells to produce cytoplasts and karyoplasts was constructed. Mouse L-cells were enucleated by exposing to cytochalasine B (10 gamma/ml) followed by centrifugation. The fraction of cells enucleated in the chamber was about 98%. The life time of cytoplasts in cultural medium after their enucleation was 48 hours (sometimes 56-72 hours) as tested by vital neutral red staining. The cytoplasts that survived were shown to accumulate large lysosomes, and the evidence of appearing ring-like fibrillar structures was provided using a simple technique of cytoskeleton observation under light microscope.
M E
ME
Neverova
L A
LA
Kirsanova
Iu S
IuS
Chentsov
D000818:Animals; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D003571:Cytochalasin B / Q000494:pharmacology*; D003584:Cytological Techniques / Q000295:instrumentation; D007739:L Cells / Q000187:drug effects / Q000648:ultrastructure; D051379:Mice; D009499:Neutral Red / Q000494:pharmacology; D013194:Staining and Labeling / Q000379:methods; D013997:Time Factors
D009499:Neutral Red; D003571:Cytochalasin B
Modifikatsiia metoda énukleatsii kletok s pomoshch'iu tsitokhalazina B i izuchenie zhiznesposobnosti tsitoplastov.