连续植块培养原代成骨细胞及其特性的比较

Culture of osteoblasts by serial explant culture and comparison of their characteristics

目的 :探讨简便、经济、高效的原代成骨细胞培养方法 ,并比较其特性。方法 :收集 1～ 4次连续植块培养获得的SD乳鼠颅盖骨原代成骨细胞 ,观察第 1～ 4次连续植块培养获得的成骨细胞在细胞形态、分裂增殖、碱性磷酸酶 (ALP)活性及骨钙素和BMP 2免疫组织化学表达的变化。结果 :连续植块培养收集的 1～ 4次原代成骨细胞 ,在细胞贴壁、变形、铺展、汇合时间 ,Gomori改良钙 -钴法ALP活性 ,骨钙素及BMP 2免疫组织化学染色表达上 ,均没有明显的区别。结论 :连续植块培养可获得与植块培养性状相同的SD乳鼠原代成骨细胞 ,且细胞数量多 ,节约经费、时间 ,操作简便。

Objective:To Investigate a simple, economical and efficient method of primary osteoblast culture and compare their characteristics. Methods:Primary osteoblasts of 1 st -4 th series explant culture from calvarial bones of neonatal Sprague Dawley rats,were collected and osteoblasts shape, mitosis, proliferation, ALP activity and immunohistological expression of osteocalcin and BMP 2 observed. Results:No difference was found in cell shape, spread, conflunce, ALP activity as well as immunohistological stain of osteocalcin and BMP 2 of the osteoblasts, which had been harvested from the 1 st -4 th series explant cultures of Sprague Dawley rats. Conclusion:Series explant culture can harvest the same kind of osteoblasts as the explant culture, and more osteoblasts can be obtained at a single time. It saves money and time and is easy to manipulate.