Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (32): 5976-5982.doi: 10.3969/j.issn.1673-8225.2010.32.019

Previous Articles     Next Articles

Migration, proliferation and differentiation of spermatogonial stem cells in vivo after transplantation

Ma Liang-hong 1,2, Ding Qiang2, Feng Li-xin3, Li Pei-jun1, Chen Fu-bao1   

  1. 1 Department of Urology, Affiliated Hospital of Ningxia Medical University, Yinchuan  750004, Ningxia Hui Autonomous Region, China; 2 Institute of Urology, Fudan University, Shanghai  200040, China; 3 Laboratory for Germline Stem Cell Research, School of Medicine, Shanghai Jiao Tong University, Shanghai  200025, China
  • Online:2010-08-06 Published:2010-08-06
  • About author:Ma Liang-hong☆, Doctor, Associate chief physician, Department of Urology, Affiliated Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Institute of Urology, Fudan University, Shanghai 200040, China
  • Supported by:

    the National Natural Science Foundation of China, No. 30860282*; the Natural Science Foundation of Ningxia Hui Autonomous Region, No.NZ08107*

PDF

358

Abstract:

BACKGROUND: Spermatogonial stem cell transplantation has potential clinical value for the treatment of male infertility. However, the process of stem cell migration, proliferation and differentiation in vivo is not yet entirely clear.
OBJECTIVE: To observe the migration, proliferation and differentiation of spermatogonial stem cells in vivo after transplantation. 
METHODS: Using C57BL/6 mice of postnatal 6-10 days as the germ cell donors, and male germ cells were obtained by combination with compound enzymatic digestions, differential velocity adherent technique and discontinuous Percoll density gradient centrifugation. Male C57BL/6 mice of postnatal 6 weeks were used as the recipients. They had been injected busulfan intraperitoneally, which could destroy endogenous spermatogenic function. In the experimental group, spermatogonial stem cells from donors were infused into the receptor testis using seminiferous tubule microinjection. We traced the PKH26-GL fluorescent-labeled cells in the recipient testes after transplantation. The migration process was observed in vivo. Western blot and RFQ-PCR were utilized to measure α6-Integrin, c-kit, SCF protein and mRNA changes in testis tissues. Normal mice without receiving busulfan treatment and cell transplantation were selected as the parallel positive control group. Mice which were microinjected media into seminiferous tubules in unilateral testes served as the parallel negative control group.
RESULTS AND CONCLUSION: Tracing the PKH26-GL fluorescent-labeled cells, partial transplanted cells immigrated into basal membrane of the seminiferous tubules at one week after transplantation. At one month after transplantation, these cells had immigrated into the basal membrane and had division growth. At 3 months, a large number of spermatid formed in the seminiferous tubule. At 1, 2 and 3 months, the expression level of α6-Integrin and c-kit protein was increased gradually in each group (P < 0.01). In negative control and experimental groups, SCF protein expression levels were increased gradually (P < 0.05). α6-Integrin, c-kit and SCF mRNA expression had increased tendency in each group (P < 0.05). Results have suggested that after high-dose chemotherapy, there are a certain number of Sertoli cells in the seminiferous epithelium. This microenvironment of spermatogenesis is not been destroyed completely. Following transplantation, spermatogonial stem cells can proliferate and differentiate in microenvironment provided by recipient Sertoli cells.

CLC Number: