新生大鼠成纤维细胞原代培养

Primary Culture Fibroblast of Newborn Rats

目的探讨新生SD大鼠成纤维细胞的培养方法。方法采用胰酶消化法和组织块贴壁法原代培养成纤维细胞。采用HE染色及波形蛋白(Vimentin)免疫细胞化学染色对培养细胞进行鉴定。结果两种方法均能培养出成纤维细胞,其中酶消化法一次性可获得较多细胞,但细胞状态较差,贴壁慢,生长慢;组织块贴壁法短时间内可长出大量成纤维细胞,一周即可传代。HE染色后倒置相差显微镜下观察细胞形态典型,免疫细胞化学染色结果显示Vimentin呈阳性反应。结论组织块贴壁培养法是获取成纤维细胞简单,可行,快速的方法。

Objective To study the method to culture fibroblast of newborn rats. Methods The primary fihro-blasts were cultured by the method of trypsin digestion and tissue adherent respectively. Hematoxylin eosin staining and vimentin immunoh-istochemical staining were used to identify fibroblast. Results The primary fibroblast can be cultured by both two methods. More fibroblasts were obtained by trypsin digestion than tissue adherent, but with poor cell state and slow adherent and growth. A large number of fibroblasts can be grown using method of tissue adherent in a short period of time, a week later, cells can be subcuhured. The passaged cells were typael fibroblast morphous under inverted phase contrast microscope and showed immunocytochemieal staining positively for Vimentin. Conclusions Adherent tissue culture is a simple, feasible and rapid method to obtain fibroblasts.