一种颈椎后纵韧带细胞的原代培养方法  

Primary culture of cervical posterior longitudinal ligament cells

作　　者：贾宽[1] 徐乐勤[1] 李晓锋[1] 王拥军[1] 周重建[1] 

机构地区：[1]上海中医药大学脊柱病研究所,上海200032

出　　处：《上海中医药杂志》2010年第2期8-10,共3页Shanghai Journal of Traditional Chinese Medicine

基　　金：上海市教委科研创新项目(08YZ56);上海市科技启明星基金项目(07QA14051);国家自然科学基金青年项目(30600829);国家杰出青年科学基金(30625043);上海市教委博士点基金(2006B10);国家自然科学基金青年基金(30801478);上海市优秀学科带头人计划(08XD1404000);国家自然科学基金青年项目(30701118)

摘　　要：目的建立颈椎后纵韧带成纤维细胞的体外培养方法。方法采用组织块培养法培养小鼠颈椎后纵韧带细胞,行HE染色,光镜及电镜观察;采用免疫细胞化学法对其进行鉴定。结果倒置相差显微镜下观察细胞呈梭形,胞质近中央处有椭圆形胞核,细胞呈放射状、漩涡状等走行形式。HE染色见细胞为长梭形,胞核为淡蓝色,胞质为粉红色。透射电镜下见胞质内有较为丰富的粗面内质网和高尔基体,线粒体呈卵圆形,常染色质丰富,异染色质靠近核膜,有核仁。免疫细胞化学鉴定表达波形蛋白,角蛋白表达为阴性。结论在体外可成功培养出颈椎后纵韧带成纤维细胞,第3-5代体外培养细胞是细胞实验的最佳选择。Objective To establish the culture of the cervical posterior longitudinal ligament cells in vitro. Methods The tissue block culture method was used to culture the mice＇s cervical posterior longitudinal ligament cells. The light and electron microscope were used to observe the cell morphology, and immunocytochemistry stain to identify the cells. Results The cells with fusiform shape and the ellipse nucleus near the center of the cytoplasm were seen by inverted phase contrast microscope. HE stain showed the cells with fusiform shape, light blue nucleus and pink cytoplasm. Transmission electron microscope observations showed the presence of affluent rough endoplasmic reticulum and golgi body and oval mitochondria in the cytoplasm, with plentiful euchromatin and the heterochromatin near the karyolemma. Immunocytochemistry stain showed the vimentin positive and the keratinose negative. Conclusion We successfully culture the cervical posterior longitudinal ligament cells in vitro; the cells between third to fifth generations are the best option for the cell experiments.

关 键 词：颈椎后纵韧带成纤维细胞 形态学 小鼠 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学] R686.5[医药卫生—基础医学]