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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 an in vitro culture method of cervical posterior longitudinal ligament fibroblasts. Methods The posterior longitudinal ligament cells of cervical spine were cultured by tissue culture method. The cells were observed with HE staining, light microscope and electron microscope. Immunocytochemistry was used to identify the cells. Results The cells were spindle-shaped under the inverted phase contrast microscope. The nucleus of the cytoplasm were oval nuclei near the center, and the cells appeared radial and swirling. HE stained cells were long fusiform, pale blue nucleus, the cytoplasm is pink. Under the transmission electron microscope, the cytoplasm was found to be rich in rough endoplasmic reticulum and Golgi apparatus. The mitochondria were oval in shape with abundant euchromatin. The heterochromatin was close to the nuclear membrane and nucleolus. Immunocytochemistry identified the expression of vimentin, which was negative for keratin expression. Conclusion The cervical posterior longitudinal ligament fibroblasts can be successfully cultivated in vitro. The 3-5th generation in vitro cultured cells are the best choice for cell experiment.