组织工程椎间盘纤维环种子细胞的培养和鉴定

目的建立兔椎间盘纤维环细胞体外培养体系，为组织工程椎间盘纤维环的研究提供种子细胞。方法采用酶消化法分离兔椎间盘纤维环细胞，进行单层培养，观察其形态结构和生物学特性，通过甲苯胺蓝、免疫细胞化学染色等方法对其细胞表型进行鉴定。结果成功进行细胞单层培养，细胞形态为圆形或多角形；原代细胞生长较慢，第一代细胞生长加快；细胞具有甲苯胺蓝异染性；有I、Ⅱ型胶原的阳性表达。结论成功培养兔椎间盘纤维环细胞，表型鉴定符合纤维环细胞特征，能够为组织工程椎间盘纤维环的研究提供种子细胞。

Culture and identification of annulus fibrosus tissue engineering seed cell in vitro

Objective To establish the culture system of the rabblit annulus fibrosus cells supplying the seed cells for annulus fibrosus disci intervertebralis tissue engineering. Methods The annulus fibrusus cells were isolated with enzyme digestion and grew as monolayer. The morpheus, structure and biological characteristic were detected; and the phenotype was evaluated with the toluiding blue staining, and immunocytochemistry means detecion. Results The cells were cultured in monolayer with the round or polygona morphology. The cells displayed intense tolniding bule metachromasia. The cells expressed collagen type I and type IT . The primary cells grew moire slowly than the first passage cells. Conclusion Rabbit annulus fibrsus cells can be cultured in vitro and it provides seed cells for annulus fibrosus disci intervertebmlis tissue engineering.