Document Detail


Techniques for purifying L-cell karyoplasts with minimal amounts of cytoplasm.
MedLine Citation:
PMID:  7444714     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Mouse L-929 cells centrifuged in the presence of cytochalasin B (CB) separate into cytoplasmic (cytoplasts) and nuclear (karyoplasts) fractions, but both of these fractions contain some whole-cell contaminants. We describe here two techniques for eliminating most of these whole cells from the karyoplast population. The first involves incubating cells in the presence of tantalum (Ta) particles, followed by enucleation and separation, on a Ficoll gradient. The small karyoplasts separate from the whole cells which contain a greater number of Ta particles. The second procedure uses a cell sorter that allows purification of karyoplasts from whole cells on the basis of light scatter. Both methods remove the whole-cell contaminants and the larger, and possibly viable, karyoplasts from the smaller karyoplast population. These purified karyoplasts are unable to regenerate but may be rescued by fusion with cytoplasts.
Authors:
M A Clark; J W Shay; L Goldstein
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Somatic cell genetics     Volume:  6     ISSN:  0098-0366     ISO Abbreviation:  Somatic Cell Genet.     Publication Date:  1980 Jul 
Date Detail:
Created Date:  1981-02-24     Completed Date:  1981-02-24     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7506054     Medline TA:  Somatic Cell Genet     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  455-64     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Fractionation / methods*
Cell Nucleus*
Colloids
Cytochalasin B
Ficoll
L Cells (Cell Line) / ultrastructure*
Mice
Tantalum
Chemical
Reg. No./Substance:
0/Colloids; 14930-96-2/Cytochalasin B; 25702-74-3/Ficoll; 7440-25-7/Tantalum

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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