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Immortalized sertoli cell lines sk11 and sk9 and binding of spermatids in vitro

Asian Journal of Andrology 2007年第3期9卷 312-320页

作者： Katja M. Wolski Caroline Feig Christiane Kirchhoff Don F. Cameron Department of Pathology and Cell Biology School of Basic Biomedical Sciences University of South Florida Collegeof Medicine Tampa Florida 33612 USA Department of Molecular Andrology University Clinics Eppendorf University of Hamburg Hamburg Germany

Aim： To determine the effectiveness of the ski 1, sk9 and ski 1 TNUA5 sertoli cell lines in binding germ cells in vitro. Methods： The immortalized sertoli cell lines sk9, ski 1 and ski 1 TNUA5 were used in co-culture experiments with germ cells in media with or without reproductive hormones and incubated for 44 h at 32℃. The number of germ cells bound to sertoli cells was then determined and statistically analyzed. Western blot analysis and reverse transcriptasepolymerase 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 ceils showed more lipid accumulation in their cytoplasm, indicating active phagocytosis. Western blot analysis in the ski I 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 ski l TNUA5 sertoli cells increased the expression of espin, as did the addition of FSH with germ cells. RT-PCR studies of the ski I TNUA5 cells indicated that the mRNA for FSH receptor decreased with successive passages. Conclusion： In vitro binding between isolated germ cells and sk9, skll or skll 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.

关键词： sk sertoli cells immortalized sertoli cells sertoli-germ cell binding sertoli-germ cell co-culture sertoli-spermatid junctionalcomplex in vitro cell-cell binding

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