[The research of xeno-lymphocytes apoptosis induced by Sertoli cells isolated by modified methods in vitro]

Zhonghua Wai Ke Za Zhi. 2007 Mar 1;45(5):331-4.
[Article in Chinese]

Abstract

Objective: To modify the isolation and culture method of Sertoli cells and investigate its' effects on xeno-lymphocytes apoptosis.

Methods: Sertoli cells which was isolated from 2 - 4 week-old Sprague Dawley (SD) rats, were successfully prepared by collagenase type V, trypsin and DNase I and then identified by electron microscope. Viability and apoptosis of cultured cells were measured by flow cytometry. The apoptosis rates of Balb/c mouse lymphocytes were examined which were co-cultured with Sertoli cells of SD rats by flow cytometry, too. The expression of FasL, TGF-beta(1) and clusterin on Sertoli cells were detected by immunocytochemistry.

Results: In the co-cultured system, Sertoli cells accounted for more than 90%. The viability of Sertoli cells was above 95% and the apoptosis rate was 10.87% +/- 3.87% in this study. The lymphocytes apoptosis ratio was 15.52% +/- 0.17% (P < 0.01). Streptavidin-biotin-peroxidase-complex immunochemistry staining showed that the Sertoli cells could express FasL, TGF-beta(1) and clusterin, respectively.

Conclusions: It indicates that the expression of FasL, TGF-beta(1) on the Sertoli cells might relate to the immune privilege, and it supposed to be benefit for xenotransplantation.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis*
  • Cell Culture Techniques / methods
  • Cell Survival / physiology
  • Cells, Cultured
  • Clusterin / metabolism
  • Coculture Techniques
  • Fas Ligand Protein / metabolism
  • Flow Cytometry
  • Immunohistochemistry
  • Lymphocytes / cytology*
  • Lymphocytes / physiology
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Microscopy, Electron, Transmission
  • Rats
  • Rats, Sprague-Dawley
  • Sertoli Cells / cytology*
  • Sertoli Cells / metabolism
  • Sertoli Cells / ultrastructure
  • Transforming Growth Factor beta / metabolism

Substances

  • Clusterin
  • Fas Ligand Protein
  • Transforming Growth Factor beta