Document Detail


Isolation of hormone responsive uterine stromal cells: an in vitro model for stromal cell proliferation and differentiation.
MedLine Citation:
PMID:  16251733     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The female sex hormones estrogen and progesterone stimulate proliferation and differentiation of human and rodent uterine cells. The purpose of this chapter is to provide a method for isolating hormone-responsive rat uterine stromal cell lines that can be used to study steroid control of the cell cycle. Uteri from ovariectomized rats are differentially digested with trypsin to separate epithelial and stromal cells. The stromal cells are cultured in a standard growth medium containing 10% fetal bovine serum. After several passages, the purity of the stromal cell lines is determined using immunocytochemistry. Cell proliferation is studied by culturing the stromal cells in serum-free medium containing sex steroids and other mitogens. Cell cycle progression is assessed by flow cytometry, 3H-thymidine and BrdU incorporation, whereas proliferation is monitored using the MTT assay. Cell cycle regulators are visualized by Northern and Western blotting whereas cyclin-cyclin-dependent kinase activity is monitored using immune complex kinase assays. Uterine stromal cell lines isolated using the methods reported in this chapter provide a suitable model system to investigate the signal transduction events that stimulate hormone-dependent control of the cell cycle.
Authors:
Virginia Rider
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Methods in molecular medicine     Volume:  121     ISSN:  1543-1894     ISO Abbreviation:  Methods Mol. Med.     Publication Date:  2006  
Date Detail:
Created Date:  2005-10-27     Completed Date:  2005-12-20     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  101123138     Medline TA:  Methods Mol Med     Country:  United States    
Other Details:
Languages:  eng     Pagination:  57-67     Citation Subset:  IM    
Affiliation:
Department of Biology, Pittsburg State University, KS, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Culture Techniques
Cell Differentiation* / drug effects
Cell Proliferation / drug effects
Cell Separation / methods*
Cells, Cultured
Estradiol / pharmacology*
Estrogen Receptor alpha / metabolism
Female
Freezing
Progesterone / pharmacology*
Rats
Stromal Cells / drug effects*
Uterus / cytology*
Chemical
Reg. No./Substance:
0/Estrogen Receptor alpha; 50-28-2/Estradiol; 57-83-0/Progesterone

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


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