支持细胞体外培养研究进展

Advances in Sertoli Cell Cultures In Vitro

总结了支持细胞 (Sertolicell,Sc)原代培养的 3种方法 :Sc分离培养、生殖细胞 Sc共培养及组织培养 (器官培养 )。采用胰蛋白酶、胶原酶及透明质酸酶等消化方法从动物睾丸分离Sc ,将纯化的Sc平铺在Matrigel包被的培养皿中作体外原代培养 ,培养基通常为 1∶1(vol/vol)无血清Ham sF12营养液和Dulbecco改良Eagle培养基 (DulbeccomodifiedEaglemedium ,DMEM) (F12 /DMEM)。培养条件 :5 %CO2 、35℃。生殖细胞 Sc共培养可模拟体内精子发生的微环境 ,用于研究生殖细胞与Sc之间的相互作用。同时也总结通过转基因动物建立的永生Sc株 。

s:Three methods of primary Sertoli cell culture were summarized. Sertoli cells were isolated from the testes by enzymatic digestion using trypsin, collagenase and hyaluronidase. Then cells were plated on Matrigel coated dishes and cultured in a humidified atmosphere of 5% CO 2 at 35℃, The medium used in primary culture was serum free Hams F12 nutrient mixture/Dulbeccos modified Eagles medium (F12/DMEM, 1∶1, vol/vol). Germ Sertoli cell coculture system, which could imitate the microenvironment of spermatogenesis, was used in study on germ Sertoli cell interactions. Permanent cell lines, which exhibited morphological and ultrastructural properties characteristic of Sertoli cell, had been established by transgenic animals.