大鼠成骨细胞的体外培养和鉴定

Identification of Rat Osteoblasts Cultured in vitro

目的探讨正常新生大鼠颅骨中分离培养大批成骨细胞并进行功能鉴定。方法将新生SD大鼠处死,无菌条件下取出颅骨,剔净后剪成1 mm×1 mm×1 mm碎块,0.1%胶原酶II消化15 min,碎骨块接种于培养瓶中培养,通过观察细胞形态学及钙化结节、碱性磷酸酶染色进行鉴定。结果用改良组织块法分离的大鼠成骨细胞,在体外培养时可维持其在体内的功能:合成碱性磷酸酶,最终能形成矿化结节。结论用改良组织块法进行大鼠成骨细胞培养,方便易行,可分离、培养大量高纯度成骨细胞,在体外传代后仍保留其表型特征。

Objective To culture and identify the functions of the rat osteoblasts （ROB） which wrere harvested from the cranial bone of normal subjects in vitro. Methods The newly born SD rats were sacrificed and the cranial bones of the rats were obtained cleanly and cut to blocks of lmm x lmm x lmm . After digested by colla- genaseII for 15 rain, the blocks were put into the culture flask. The phenotype, mineralized nodes and the expression of alkaline phosphatase were studied with alizarin red - S staining and modified Gomori＇ s assays, respectively. Results The ROB cells purified by modified method of tissue block functioned normally in vitro as observed in vivo. They showed the osteoblastic phenotypes such as their synthesis of alkaline phosphatase and osteocalcin and formatian of mineralized nodes. Conclusion The study indicates that it is easy to purify plentiful ROBs with normal functions by modified method of tissue block, which can be used in the next research of bone metabolism