用多次酶消化法体外培养的大鼠成骨细胞及其鉴定

目的 针对目前体外培养成骨细胞方法的不足,建立一种理想的原代培养成骨细胞的方法,为骨替代材料的研究提供成骨细胞.方法 本实验用新生1～2天大鼠颅盖骨,采用多次酶消化法进行细胞体外培养.倒置显微镜观察细胞形态,并对其碱性磷酸酶（ALP）活性及矿化能力进行鉴定.结果 所培养细胞具有成骨细胞的形态学特征及体内成骨细胞的生物学行为.结论 本实验体外培养成骨细胞的方法切实可行,可为骨替代材料的研究提供种子细胞,也可为骨细胞生物学和骨组织工程的研究提供一种客观而有效的实验手段.

Culture in Vitro by Multiple Enzyme Digestion and Identification of Rat Osteoblasts

Objective Based on the advantages and disadvantages of technique commonly for isolating osteoblasts from bone tissue, an ideal method of primary culture osteoblast was established to provide the study on bone substitutes with osteoblasts. Methods The bone tissue from the calvaria of 1 - 2day rats was digested many times to isolate and collect cells. The harvested cells were cultured in vitro. The identification was conducted by cell morphology, histochemical stain of alkaline phosphatase (ALP) and calcified nodules stain. Results The cultured cells possessed the morphological characteristic and biological behavior of osteoblasts. Conclusion This method of primary culture osteoblasts is practicable, efficient and effective, and the cultured osteoblasts can be applied to related studies in vitro.