Document Detail


Characterization and functionality of proliferative human Sertoli cells.
MedLine Citation:
PMID:  21054948     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
It has long been thought that mammalian Sertoli cells are terminally differentiated and nondividing postpuberty. For most previous in vitro studies immature rodent testes have been the source of Sertoli cells and these have shown little proliferative ability when cultured. We have isolated and characterized Sertoli cells from human cadaveric testes from seven donors ranging from 12 to 36 years of age. The cells proliferated readily in vitro under the optimized conditions used with a doubling time of approximately 4 days. Nuclear 5-ethynyl-2'-deoxyuridine (EdU) incorporation confirmed that dividing cells represented the majority of the population. Classical Sertoli cell ultrastructural features, lipid droplet accumulation, and immunoexpression of GATA-4, Sox9, and the FSH receptor (FSHr) were observed by electron and fluorescence microscopy, respectively. Flow cytometry revealed the expression of GATA-4 and Sox9 by more than 99% of the cells, and abundant expression of a number of markers indicative of multipotent mesenchymal cells. Low detection of endogenous alkaline phosphatase activity after passaging showed that few peritubular myoid cells were present. GATA-4 and SOX9 expression were confirmed by reverse transcription polymerase chain reaction (RT-PCR), along with expression of stem cell factor (SCF), glial cell line-derived neurotrophic factor (GDNF), and bone morphogenic protein 4 (BMP4). Tight junctions were formed by Sertoli cells plated on transwell inserts coated with fibronectin as revealed by increased transepithelial electrical resistance (TER) and polarized secretion of the immunoregulatory protein, galectin-1. These primary Sertoli cell populations could be expanded dramatically in vitro and could be cryopreserved. The results show that functional human Sertoli cells can be propagated in vitro from testicular cells isolated from adult testis. The proliferative human Sertoli cells should have important applications in studying infertility, reproductive toxicology, testicular cancer, and spermatogenesis, and due to their unique biological properties potentially could be useful in cell therapy.
Authors:
Kitty Chui; Alpa Trivedi; C Yan Cheng; Diana B Cherbavaz; Paul F Dazin; Ai Lam Thu Huynh; James B Mitchell; Gabriel A Rabinovich; Linda J Noble-Haeusslein; Constance M John
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2010-11-05
Journal Detail:
Title:  Cell transplantation     Volume:  20     ISSN:  1555-3892     ISO Abbreviation:  Cell Transplant     Publication Date:  2011  
Date Detail:
Created Date:  2011-10-19     Completed Date:  2012-02-09     Revised Date:  2014-07-15    
Medline Journal Info:
Nlm Unique ID:  9208854     Medline TA:  Cell Transplant     Country:  United States    
Other Details:
Languages:  eng     Pagination:  619-35     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Adolescent
Adult
Bone Morphogenetic Protein 4 / genetics,  metabolism
Cell Proliferation
Child
Deoxyuridine / analogs & derivatives,  pharmacology
GATA4 Transcription Factor / genetics,  metabolism
Galectin 1 / metabolism
Glial Cell Line-Derived Neurotrophic Factor / genetics,  metabolism
Humans
Male
Receptors, FSH / metabolism
SOX9 Transcription Factor / genetics,  metabolism
Sertoli Cells / cytology*,  metabolism,  ultrastructure
Stem Cell Factor / genetics,  metabolism
Grant Support
ID/Acronym/Agency:
1R43HD056620/HD/NICHD NIH HHS; 2 U42 RR006042-13/RR/NCRR NIH HHS; 2R44HD042333/HD/NICHD NIH HHS; 5R01HD056034/HD/NICHD NIH HHS; R01 HD056034/HD/NICHD NIH HHS; R43 HD056620/HD/NICHD NIH HHS; R44 HD042333/HD/NICHD NIH HHS; U42 RR006042/RR/NCRR NIH HHS
Chemical
Reg. No./Substance:
0/BMP4 protein, human; 0/Bone Morphogenetic Protein 4; 0/GATA4 Transcription Factor; 0/Galectin 1; 0/Glial Cell Line-Derived Neurotrophic Factor; 0/Receptors, FSH; 0/SOX9 Transcription Factor; 0/SOX9 protein, human; 0/Stem Cell Factor; 61135-33-9/5-ethynyl-2'-deoxyuridine; W78I7AY22C/Deoxyuridine

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


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