摘要
目的：建立乳鼠成骨细胞体外培养方法，探讨该方法的可行性和应用价值。方法：用出生1～3d乳鼠颅骨,采用多次胶原酶消化法进行细胞体外培养。倒置显微镜观察细胞形态，对其碱性磷酸酶（ALP）活性及矿化能力进行鉴定，并测定细胞生长曲线。结果：原代培养24h后,大量细胞贴壁生长．细胞呈圆形，48h后,贴壁细胞呈长梭形、三角形或不规则多边形．并且贴壁细胞伸出2～3个突起,胞质透亮、饱满，7d后细胞铺满整个平皿底面。经鉴定，培养细胞具有体内成骨细胞的生物学特性。细胞接种后第1与第2个24h为细胞的潜伏适应期，第3与第7个24h生长曲线基本为线性曲线，是细胞的对数生长期。结论：采用胶原酶消化法分离培养成骨细胞的方法切实可行。
Objective To establish an experimental model of osteoblasts of newborn mouse in vitro, a procedure of osteoblast isolation and culture was developed, and the feasibility of this technique and its practical value were investigated. Methods Osteoblasts were isolated from fetal mice within 1 - 3 day. Collagenase digestion of the skull of fetal mice was conducted repeatedly, and the collected cells were cultured in vitro. The osteoblasts were identified by cell morphology, histochemical alkaline phosphatase （A1Pase） staining and calcified nodules staining. The growth curve was recorded. Results Many cells showed adherence to the culture plat after 24 hours of cell culture. The cells were of round appearance. After 48 hours of cell culture, the appearance of cells became long fusiform, triangular or irregular polygonal and often had 2 or 3 projections. After 7 days, the cells grew in full flat plate undersurface. The cultured cells possessed biological behavior of osteoblasts. The growth curve showed that the first and second 24 hours were the adaptive phase, the third to the seventh 24 hours was log phase growth. Conclusion Repeatedly digestion of skull of fetal mice and culture of osteoblasts in vitro is a practical, efficient and effective method.
出处
《中国实验动物学报》
CAS
CSCD
2007年第3期231-233,I0006,共4页
Acta Laboratorium Animalis Scientia Sinica
基金
国家自然科学基金资助项目(编号:30560131)
关键词
成骨细胞
胶原酶
原代培养
Osteoblast
Primary culture
Collagenase
Mouse, fetal
