组织块法培养SD大鼠的成骨细胞及鉴定

背景：获得足够量高纯度的成骨细胞比较困难,因此掌握简单而快速提取成骨细胞并进行培养鉴定势在必行。目的：应用组织块法对SD大鼠成骨细胞的体外培养与鉴定。方法：取新生(<24 h)SD大鼠颅骨,去除周围多余的组织,将剔净颅骨剪成1 mm3大小的碎块,分别用常规方法和组织块法(改良)方法进行成骨细胞培养,从形态学、碱性磷酸酶和茜苏红染色等方法鉴定。结果与结论：利用改良方法培养的细胞具有典型的成骨细胞形态特征,碱性磷酸酶呈阳性染色,茜素红染色后有矿化结节的形成。组织块法培养的成骨细胞具有典型的成骨细胞成分单一、细胞培养时间短、数量、纯度、密度均一致,从而的得到稳定的成骨细胞系,为进行体外实验建立了良好的平台。

Explant culture and identification of SD rat’s osteoblasts

BACKGROUND:It is difficult to acquire sufficient osteoblasts with high purity. Therefore, it is imperative to master simple and fast extraction osteoblast method for culture and identification.OBJECTIVE:To culture in vitro and identify SD rat’s osteoblasts with explant culture.METHODS:The newborn (< 24 hours) SD rats were executed, and the skulls were taken out. Excess tissues around the skull were removed. The picked skull was cut off into 1 mm3 pieces. The osteoblasts were cultured by conventional method and tissue explant (modified method) as well as identified with morphology, alkaline phosphatase assay and Von Kossa staining methods.RESULTS AND CONCLUSION:The osteoblasts cultured by tissue explant had typical morphological characteristics; alkaline phosphatase was positive staining, and mineralization nodules formed after Von Kossa staining. Besides, the osteoblasts had unitary cell component which was typical, and the culture time was short, what’s more, the quantity, purity and density were coincident. These findings suggest that the osteoblast lineage acquired is stable, which provides a good plate for in vitro experiments.