【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。更多还原

【Abstract】 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.更多还原