成年大鼠成骨细胞体外培养及细胞凋亡的观察

Observation on the external cultivate and cell-wither of mature rat osteoblasts

目的 为研究骨重建过程中成骨细胞的关键作用 ,建立成年大鼠成骨细胞体外培养方法并进行细胞凋亡的观察。方法 1 5周龄SD大鼠 ,在无菌条件下取颅盖骨置于培养瓶中DMEM培养液 ,内含体积分数为 1 0 %的胎牛血清 (FCS) ,在体积分数为 5 %的CO2 、37℃孵箱内培养。按原位DNA末端标记 (TUNEL)检测试剂盒方法染色 ,以荧光显微镜观察凋亡。结果 经活体观察、ABC法Ⅰ型胶原染色和碱性酸酶染色 ,显示培养细胞为成骨细胞。

Objective To study the key effect of osteoblasts in the process of bone reduilding,and to set up a way to study external cultivate and cell wither of mature rat osteoblasts.Methods Obtained the cranial bones after executed rats and cultivated in vitro in conditional DMEM medium with 10% of fetal boveine serum,hatched under 37℃ and 5% of CO 2.Then observed the cell wither of osteoblasts under fluorescent light microscope by signed situs DNA tip after stained with TUNEL.Results The cultivated cells were conformed to be osteoblasts under microscope by type Ⅰ fibrogen stained by ABC method,and stained alkaline phosphorylase.Conclusion The experiment succeeded to set up a way to study the external cultivated and cell wither of mature rat.