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目的探讨从不同年龄的人获得黄韧带组织与原代培养细胞游出之间的关系,以提高黄韧带细胞原代培养的成功率。方法采用酶消化加组织块培养法,对来自21例不同年龄腰椎管狭窄和腰椎间盘突出症患者的黄韧带组织进行黄韧带细胞原代培养,倒置相差显微镜下观察细胞从组织块迁出时间、形态和生长状态;采用细胞免疫荧光染色法对传第3代细胞进行Ⅰ型胶原和波形蛋白表达鉴定;对组织块内细胞游出率和年龄进行相关性分析。结果倒置显微镜下见黄韧带细胞形态生长良好,培养细胞的波形蛋白和Ⅰ型胶原免疫荧光化学染色均呈阳性。患者年龄与黄韧带组织块原代培养时细胞游出率之间呈负相关关系(r=-0.618,P<0.05),年轻患者黄韧带组织细胞游出率高。结论原代培养人黄韧带细胞时尽量采用年轻人黄韧带组织。
OBJECTIVE: To investigate the relationship between the ligamentum flavum (LPL) and the primary cultured cells derived from different age groups in order to improve the success rate of primary ligament cells culture. Methods The ligament vulgaris tissue culture method was used to culture the ligamentum flavum cell from the ligamentum flavum of 21 patients with lumbar spinal canal stenosis and lumbar disc herniation at different ages. The cells were removed from the tissue block under inverted phase contrast microscope. Morphology and growth status. The expression of type I collagen and vimentin were detected by immunofluorescence staining. The correlation between cell migration rate and age was analyzed. Results The cells of the ligamentum flavum grew well under the inverted microscope, and the vimentin and type Ⅰ collagen immunofluorescence staining were all positive. There was a negative correlation between the age of patients and the rate of cell migration in the primary culture of ligamentum flavum (r = -0.618, P <0.05), and the rate of lobulation of ligamentum flavum in young patients was high. Conclusion The primary culture of human ligamentum flavum cells try to use young yellow ligament tissue.