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目的探讨建立乳鼠成骨细胞体外培养方法的可行性及成骨细胞的生物学特性。方法取出生后24h内乳鼠颅盖骨,采用多次复合胶原酶消化法进行消化及体外培养,通过倒置相差显微镜观察细胞形态,进行成骨细胞HE染色,碱性磷酸酶染色,基质钙结节染色及Ⅰ型胶原蛋白免疫组织化学染色,对大鼠成骨细胞进行鉴定。结果所培养细胞为成骨细胞,并生长良好,HE、碱性磷酸酶染色,钙结节染色及Ⅰ型胶原蛋白染色均阳性。结论采用复合胶原酶消化法分离培养的成骨细胞具有典型成骨细胞生物学特性,且成分较单一。
Objective To investigate the feasibility of establishing osteoblasts cultured in vitro and the biological characteristics of osteoblasts. Methods The calvaria of neonatal rats were taken out within 24 hours after birth. The cells were digested with compound collagenase digestion and cultured in vitro. The morphological changes of cells were observed by inverted phase contrast microscope. HE staining, alkaline phosphatase staining, Day staining and type Ⅰ collagen immunohistochemical staining of rat osteoblasts were identified. Results The cultured cells were osteoblasts and grew well. HE, ALP staining, calcium nodule staining and type Ⅰ collagen staining were all positive. Conclusion The osteoblasts isolated and cultured by compound collagenase digestion have the typical characteristics of osteoblasts, and the components are single.