周细胞的原代培养

目的:探讨Wistar大鼠视网膜毛细血管周细胞(pericyte,PC)的原代培养和鉴定方法。方法:结合视网膜微血管的消化分离,采用含10%胎牛血清的DMEM培养基选择性培养PC,通过活细胞观察原代PC的形态、生长特性以及与血管碎片之间的关系,同时应用免疫细胞化学染色来鉴定PC。结果:选择性培养获得的PC的纯度达到95%以上,并能连续传代。该细胞呈长梭形或星芒状,漩涡或栅栏状生长,无接触性抑制,单核,偶见双核,核卵圆形,细胞浆丰富,α-SMA、PDGFR-β染色阳性。结论:通过对视网膜微血管的消化分离能够获得较为纯净的PC。

The Primary Culture of pericytes

Objectives: To cultivate purify and identify pericytes derived from rat retinal capillaries.Methods: We combined the methods of digestion and distinguish of retinal capillary,used Dulbecco’s modified Eagle’s medium(DMEM) containing 20% fetal bovine serum,rat retinal capillary pericytes were cultured in vitro with digesting culture method of retinal microvasculature,and the cultured cells were identified by immunohistochemical staining.The morphous of pericytes,growth character and relationship between vascular fragment and the cells cultured in vitro were observed by using inverted microscope.Results: The purity of the cells cultured in vitro was higher than 95%,and the cells could be continued serially.The shape of pericyte was long fusiform,asterism,whirlpool or palisade,no restraint with contact,mononuclear occasional dikaryon,nucleus ovoid,cytoplasm rich,α-SMA and PDGFR-β were positive.Conclusion:Higher purity retinal capillary PC cultured in vitro can be obtained through the selective culture method.