新生小鼠成骨细胞原代培养方法的实验研究

目的 验证及比较成骨细胞原代培养常用方法,探讨两种原代培养方法的可行性和应用价值.方法 取出生后24 h昆明小鼠乳鼠顶骨,以含血清胶原酶消化法和分段胶原酶消化法分离成骨细胞,进行细胞形态观察,以噻唑蓝(MTT)比色实验法绘制生长曲线,以台盼兰排斥法计数活细胞率.结果 与含血清胶原酶消化法相比,分段胶原酶消化法提取的成骨细胞产量多;细胞存活率高(P<0.01);对细胞损伤小,细胞纯度高.消化时问易于控制.结论 分段胶原酶消化法是一种方便、高效、细胞生物学特性稳定的原代成骨细胞分离培养方法.

Abstract：

Objective To confirm an alternative method for primary culture of osteoblasts through comparison of two primary culture methods. Methods Calvarias were dissected from newborn Kunming mice, osteoblasts were isolated with serum-containing collagenase digestion method and sequential digestion method respectively. The osteoblasts were observed under invert microscope, the growth curve was made with the application of MTT method, the rate of living osteoblasts was counted with trypan blue method respectively. Results Compared with the serum-containing collagenase digestion method, the sequential digestion method presented higher production of osteoblasts, higher cell survival rate (P<0.01), little damage on osteoblasts and easier control of the digestion time. Conclusion The sequential digestion of calvaria method is simple, efficient and stable for primary osteoblasts culture.

Primary Culture of Osteoblasts of Neonatal Mice: a Comparative Research

Objective To confirm an alternative method for primary culture of osteoblasts through comparison of two primary culture methods. Methods Calvarias were dissected from newborn Kunming mice, osteoblasts were isolated with serum-containing collagenase digestion method and sequential digestion method respectively. The osteoblasts were observed under invert microscope, the growth curve was made with the application of MTT method, the rate of living osteoblasts was counted with trypan blue method respectively. Results Compared with the serum-containing collagenase digestion method, the sequential digestion method presented higher production of osteoblasts, higher cell survival rate (P<0.01), little damage on osteoblasts and easier control of the digestion time. Conclusion The sequential digestion of calvaria method is simple, efficient and stable for primary osteoblasts culture.