软骨终板细胞生物学特性及体外退行性变的机制研究  

The biological characteristics of cartilage endplate cells and the mechanism of cartilage endplate degeneration in vitro

作　　者：刘斌[1] 瞿东滨[1] 金大地[1] 王岩松[2] 

机构地区：[1]解放军第一军医大学南方医院脊柱骨病科,广东省广州市510515 [2]哈尔滨医科大学附属第二医院脊柱外科,黑龙江省哈尔滨市150086

出　　处：《中国临床康复》2003年第29期3960-3961,F003,共3页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金资助项目(30000168);广东省自然科学基金资助项目(001105)~~

摘　　要：目的:通过建立猪腰椎间盘软骨终板细胞的培养模型,观察其演变,探讨软骨终板细胞的生物学行为及其影响因素。方法:取长枫杂交猪腰椎间盘软骨终板细胞,于200mL/L胎牛血清的杜尔伯克改良基础培养基(Dulbecco'smodifiedEagle'smedium,DMEM)中培养,建立体外软骨终板细胞培养模型。以光镜及苏木精-伊红染色观察其生物学表现;以甲苯胺蓝、番红O、Ⅱ型胶原免疫细胞化学染色对软骨终板细胞进行鉴定;观察冻存后软骨终板细胞的生物学特性。结果:原代细胞生物学性状最接近体内细胞,多次传代后细胞呈现衰老现象。经染色证实此细胞具有Ⅱ型胶原及蛋白多糖的表达。冻存细胞复苏后,细胞形态及其分化、增殖能力与冻存前基本相同。结论:猪腰椎软骨终板细胞的体外培养模型的建立,可为体外研究软骨终板的退行性变机制提供实验基础。AIM:To establish a cartilage endplate cells culture method in vitro so that we can observe the continuous changes and explore the biological action and affected factors of it. MEHTODS:The cells were isolated and harvested from the cartilage endplate of the porcine lumbar,and we cultured the cells in DMEM with 20 mL/L fetal bovine serum (FBS).Their biological phenomena was observed by light microscope and HE staining.Their biological characteristics were identified by toluidine blue and safranine O staining for proteoglycan and immunocytochemistry for type Ⅱ collagen.We also observed the changes of biological characteristics of the cells after frozen storage.RESULTS:The biological characteristics of the primary cells were the most similar to those in vivo. There was a senilism in these cells after many times passage. We could identify the cells' phenotype by staining, and frozen storage didn't change the cells' biological characteristics.

关 键 词：软骨终板 细胞生物学 体外退行性变 细胞培养 

分 类 号：R336[医药卫生—人体生理学]