Document Detail


Cell cycle synchronization of porcine granulosa cells in G1 stage with mimosine.
MedLine Citation:
PMID:  12695057     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The success of somatic cell nuclear transfer depends critically on the cell cycle stage of the donor nucleus and the recipient cytoplast. Karyoplasts in the G0 or G1 stages are considered to be the most suitable for nuclear transfer. In the present study, we used a reversible cell cycle inhibitor, mimosine, to synchronize porcine granulosa cells (GCs) in G1 phase of the cell cycle. Porcine GCs were obtained from 3 to 5mm ovarian follicles of slaughtered gilts. The effect of mimosine on the proliferation, DNA synthesis and cell cycle stage of cultured cells was examined by incorporation of radiochemical 3H-thymidine, immunocytochemical detection of incorporated thymidine analogue 5-bromo-2-deoxyuridine (BrdU) and flow cytometry analyses. Mimosine treatment of pig GCs for 24h resulted in proliferation arrest in vitro. Treatment with 0.5mM mimosine significantly (P<0.05) inhibited 3H-thymidine incorporation after 24h of culture (4.6% +/- 0.1) and after 24h of culture in serum deprived medium (41.3% +/- 3.8), in comparison to controls (100%). Inhibition of DNA synthesis was further confirmed by immunocytochemical and flow cytometry analyses. Compared with controls (78.2%), mimosine treatment for 24h increased the proportion of G0/G1 cells in the culture (85.7%) more effectively than serum starvation (SS; 81.2%). Mimosine-caused G1 arrest of porcine GCs was fully reversible and cells continued to proliferate after removing the drug, especially when they were stimulated by EGF.
Authors:
I Vacková; M Engelová; I Marinov; M Tománek
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Animal reproduction science     Volume:  77     ISSN:  0378-4320     ISO Abbreviation:  Anim. Reprod. Sci.     Publication Date:  2003 Jul 
Date Detail:
Created Date:  2003-04-15     Completed Date:  2003-08-26     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7807205     Medline TA:  Anim Reprod Sci     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  235-45     Citation Subset:  IM    
Affiliation:
Department of Biology of Reproduction, Research Institute of Animal Production, Prátelství 815, P.O. Box 1, 104 01 Prague, Uhríneves, Czech Republic.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Cycle / drug effects*
Cell Division / drug effects
Cells, Cultured
DNA / biosynthesis
Dose-Response Relationship, Drug
Epidermal Growth Factor / pharmacology
Female
Flow Cytometry
G1 Phase*
Granulosa Cells / cytology*,  drug effects
Immunohistochemistry
Mimosine / administration & dosage,  pharmacology*
Swine*
Time Factors
Chemical
Reg. No./Substance:
500-44-7/Mimosine; 62229-50-9/Epidermal Growth Factor; 9007-49-2/DNA

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


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