退变兔椎间盘软骨终板细胞培养及其形态学和生物学特征  

The biological characteristic and degeneration morphology of the rabbit endplate cartilage cell culture

作　　者：屈开钛[1] 徐永清[1] 王非[2] 

机构地区：[1]成都军区昆明总医院骨科,云南昆明650032 [2]广州军区总医院骨科,广东广州510010

出　　处：《颈腰痛杂志》2008年第5期403-407,共5页The Journal of Cervicodynia and Lumbodynia

基　　金：国家自然科学资助(编号:30400456(青年基金面上项目))

摘　　要：目的观察研究兔椎间盘软骨终板的细胞形态及表征,利用正常动物的体细胞传代培养发生功能退变的原理,制作软骨终板细胞体外培养的退变模型。方法酶消化法培养兔椎间盘软骨终板细胞,并用HE和甲苯蓝染色,进行光镜观察;使用免疫组织化学法检测特征性Ⅱ型胶原表达情况,与同种动物不同部位的软骨细胞做对照。结果镜下观察,HE染色见细胞呈多角形,核嗜碱性,圆形或椭圆形,有时见双核,胞浆内可见空泡,有弱嗜酸性。甲苯胺蓝染色可见胞浆有紫红色和红色异染出现。原代细胞与传代细胞之间,形态结构无明显差异,但多次传代后(4代),细胞则逐渐失去其原有形态,变为短梭型细胞,失去分化特性,表达Ⅱ型胶原的量逐渐减少。软骨关节细胞与终板细胞形态学相似。结论本研究提示(1)软骨终板细胞可表达其特征性产物蛋白多糖和Ⅱ型胶原;(2)软骨终板细胞在传代培养时有逐渐退变,失去分化特性的特点;(3)在应用平面培养体系时应选用前3代细胞进行实验。以上几点均为椎间盘软骨终板退变机理研究提供了细胞学基础。Objective To observe the cell morphous and exosyndrome on the rabbit endplate cartilage cell culture,to utilize the degeneration developed during normal animal body cell serial subcuhivation,it, cartilage end plate cell in vitro culture degeneration model. Methods Enzyme digestion cuhivante the rabbit endplate cartilage cell,to observe after HE stain or blue cresyl under light microscope. To use the immunohistochemical methods detect the mark of collagen Ⅱ express. Then compare with the homogeneity from different body cartilage cell as a check. Results Observe under the light microscope,HE stain found the cartilage end plate cell was shaped as polygon,the nuclear is basophilia,round or ellipse,the intracytoplasm was weak acidophilia,vacuole could be see. TB found prunosus and red metachromatie in endochylema. The primary cell and the passage cell had no difference on morphous,but after high passage （4 generations）the cell could lose the shape at very beginning gradually,shaped as short shuttle and the cell differentiation to be missing,the mass of collagen Ⅱ express also declining gradually. The cartilage joint taken as a check has the same cell morphology changes in comparation. Conclusion （1）Cartilage end plate cell can express the mark products such as proteoglycan and collagen Ⅲ. （2）Cartilage end plate cell had the characteristic of dedifferentiation during serial subcuhivation and degenerate gradually. （3）The cell to be chosen for plane cuhivante experimentize should be less than 3rd generation. All those points above aimed to provide the cytology foundation for the research on the degeneration of IVD cartilage endplate.

关 键 词：退变 软骨 细胞培养 免疫组织化学 

分 类 号：R681.53[医药卫生—骨科学] R392.11[医药卫生—外科学]