Document Detail


Immortalized Sertoli cell lines sk11 and sk9 and binding of spermatids in vitro.
MedLine Citation:
PMID:  17486271     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
AIM: To determine the effectiveness of the sk11, sk9 and sk11 TNUA5 Sertoli cell lines in binding germ cells in vitro. METHODS: The immortalized Sertoli cell lines sk9, sk11 and sk11 TNUA5 were used in co-culture experiments with germ cells in media with or without reproductive hormones and incubated for 44 h at 32 degrees . The number of germ cells bound to Sertoli cells was then determined and statistically analyzed. Western blot analysis and reverse transcriptase-polymerase chain reaction (RT-PCR) studies were employed to investigate the presence of cell adhesion proteins and follicle stimulating hormone (FSH) receptor, respectively. RESULTS: No statistical difference between the number of bound step-8 spermatids and bound pre-step 8 spermatids on Sertoli cells from any of the cell lines existed. After the addition of germ cells, Sertoli cells showed more lipid accumulation in their cytoplasm, indicating active phagocytosis. Western blot analysis in the sk11 TNUA5 line indicated the expression of N-cadherin. FSH-only and testosterone-only treatments increased N-cadherin expression, regardless of germ cell addition. The addition of germ cells to the sk11 TNUA5 Sertoli cells increased the expression of espin, as did the addition of FSH with germ cells. RT-PCR studies of the sk11 TNUA5 cells indicated that the mRNA for FSH receptor decreased with successive passages. CONCLUSION: In vitro binding between isolated germ cells and sk9, sk11 or sk11 TNUA5 Sertoli cells is not feasible, and therefore these cell lines are not useful for the in vitro investigation of Sertoli-germ cell interactions and primary Sertoli cell isolates must still be used.
Authors:
Katja M Wolski; Caroline Feig; Christiane Kirchhoff; Don F Cameron
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Asian journal of andrology     Volume:  9     ISSN:  1008-682X     ISO Abbreviation:  Asian J. Androl.     Publication Date:  2007 May 
Date Detail:
Created Date:  2007-05-08     Completed Date:  2007-07-03     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100942132     Medline TA:  Asian J Androl     Country:  China    
Other Details:
Languages:  eng     Pagination:  312-20     Citation Subset:  IM    
Affiliation:
Department of Pathology and Cell Biology, University of South Florida, College of Medicine, Tampa, FL 33612, USA. katjawolski@gmail.com
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MeSH Terms
Descriptor/Qualifier:
Animals
Cadherins / genetics,  metabolism
Cell Adhesion / physiology
Cell Line, Transformed / cytology*,  metabolism
Coculture Techniques
Gene Expression
Image Processing, Computer-Assisted
Male
Mice
Microfilament Proteins / genetics,  metabolism
RNA, Messenger / metabolism
Receptors, FSH / genetics,  metabolism
Reverse Transcriptase Polymerase Chain Reaction
Sertoli Cells / cytology*,  metabolism
Spermatids / cytology*,  metabolism
Chemical
Reg. No./Substance:
0/Cadherins; 0/Cdh2 protein, mouse; 0/Microfilament Proteins; 0/RNA, Messenger; 0/Receptors, FSH; 0/espin protein, mouse

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