Click for new scientific resources and news about Corona[COVID-19]

Paper Information

Journal:   INTERNATIONAL JOURNAL OF REPRODUCTIVE BIOMEDICINE (IRANIAN JOURNAL OF REPRODUCTIVE MEDICINE)   SPRING 2011 , Volume 9 , Number SUPPL 2; Page(s) 51 To 52.
 
Paper: 

ISOLATION AND CULTURE OF HUMAN SPERMATOGONIAL STEM CELL FROM TESTIS BIOPSY DETERMINED BY GPR125

 
 
Author(s):  GOHARBAKHSH L.*, MOHAZZAB A., SALEHKHOU SH., HEIDARI M., ZARNANI A.H., PARIVAR K., AKHOONDI M.M.
 
* DEPARTMENT OF BIOLOGY, FACULTY OF SCIENCES, ISLAMIC AZAD UNIVERSITY, SCIENCES AND RESEARCH CAMPUS, TEHRAN, IRAN
 
Abstract: 
Introduction: A high-dose of chemo and radiotherapy in cancer patients can cause infertility by damaging spermatogenesis process. This process is based on self-renewal and differentiation of a rare population of the testicular cells called spermatogonial stem cells (SSCs). Stemness of SSCs is only preserved in a specific microenvironment called SSC niches. Because of unknown signaling pathway of this niche, in vitro culture of SSCs is very difficult.
Isolation and culture of human SSCs in vitro would identify this environment in order to preserve fertility. Isolation and specific culture of human SSCs from testicular cell population in serum free media may enrich these rare cells.
Materials and Methods: Human SSCs were isolated and purified from testicular biopsies. Two enzymatic digestion steps were performed by collagenase IV and then trypsin on each biopsy consecutively. In the first 24 hrs, cells were suspended in DMEM-F12 with 10% FBS in plastic dishes. The media was changed to serum-free medium with added growth factors: human GDNF, bFGF, EGF and LIF. Colonies were passaged every 7-10 days to expand SSCs and then used antiGPR125 (a surface marker of SSCs) to determine SSCs in the culture by the immunocytochemistry test (LSAB).
Results: SSC colonies were appeared within 10 days.
They increased in numbers after 52 days. Myofibroblasts and other peritubular cells were survived without any notable proliferation. In immunocytochemistry, approximately 70-80% of cells were GPR125 positive.
Conclusion: In contrast to the other SSCs culture methods, this system is based on the testicular biopsies (against large samples) which preserves the somatic cells as a supporter. However, without extrinsic feeder cells or biomatrix, proliferation of SSCs cells was slighter than systems supported by laminin. Furthermore, these SFM was based on common DMEM-F12 instead of highly purified STEM-Pro34. Other properties of this culture will be investigated in the future.
 
Keyword(s): HUMAN SPERMATOGONIAL STEM CELL, CULTURE, ISOLATION, GPR125, INFERTILITY
 
References: 
  • ندارد
 
  Yearly Visit 42
 
Latest on Blog
Enter SID Blog