Long-term Culture and Spermatogenesis Analysis of Sertoli Cell/Germ Cell Co-culture System from Seminiferous Tubules of Rat Testis

Abstract: The development of in vitro models of testicular reproductive system may provide important tools for investigating specific mechanisms of spermatogenesis and toxicity in the testis. Although various systems have been reported， their application in reproductive studies has been limited by the poor ability to replicate the complex biochemical， molecular， and functional interactions observed in the testis. In the present study， we evaluated a significantly improved sertoli cell/germ cell co-culture (SGC) system that the proliferation and differentiation of testicle germ cells of the rat could be maintained for long-term in vitro， and spermatids produced ceaselessly. The germ cells and sertoli cells came from pieces of the testis seminiferous tubules of the rat. The co-cultured cells survived in vitro for over two months without extra growthfactor. Furthermore， RT-PCR analysed cells stabilization of cdh1、scp3、tnp2 expression confirmed the integrity of this co-culture system. Immunocytochemistry displayed the CDH1、PLZF、SCP3、SOX9 positive cells were existed in system. All the results proved that the SGC system included the spermatogonial stem cell， spermatocyte， round spermatid and sertoli cell. We concluded that this system would provide investigators with a simple and efficient alternative for the assessment of molecular mechanisms associated with both male reproduction development and reproductive toxicity.

CSTR: 32200.14.cjcb.2012.05.0007