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目的:获取较纯的大鼠(SD)胎鼠成骨细胞,为生物学评价奠定基础。方法:从SD临产孕鼠腹中取出胎鼠,机械剥离其颅骨片,酶消化法与组织块法结合培养原代混合细胞,采用差速贴壁法对成骨细胞进行纯化,不同染色法进行鉴定。结果:原代培养的细胞为混杂细胞,经2~3次纯化可完全去除纤维样细胞。成骨细胞呈三角形、多边形,碱性磷酸酶和钙结节染色阳性。结论:实验获取较纯的大鼠胎鼠成骨细胞,符合体外细胞生物学评价试验中对细胞的要求。
OBJECTIVE: To obtain more pure rat (SD) fetal rat osteoblasts and lay a foundation for biological evaluation. METHODS: Fetal rats were removed from the abdomen of SD pregnant rats and their skulls were dissected mechanically. Enzyme digestion was combined with tissue culture to culture primary mixed cells. The osteoblasts were purified by differential adherence method. Different staining methods were performed Identification. Results: The primary cultured cells were mixed cells. After 2 or 3 times of purification, fibroblast-like cells could be completely removed. Osteoblasts were triangular, polygonal, alkaline phosphatase and positive staining for calcium nodules. CONCLUSION: The pure rat fetal rat osteoblasts were obtained experimentally, which accords with the requirement of cells in vitro cell biology evaluation test.