退变兔髓核细胞和正常髓核细胞生物学特性的比较

Comparison of biological characteristics between primary and degenerative rabbit nucleus pulposus cells

目的探讨退变和正常兔髓核细胞的不同生物学特性。方法分离和培养原代兔椎间盘髓核细胞,体外传至第5代使其发生退变,细胞分成原代髓核细胞组(原代)和第5代退变细胞组(退变),用倒置显微镜和HE染色观察其形态学变化,用番红O染色观察糖胺聚糖的变化,用Ⅱ型胶原免疫组织化学观察Ⅱ型胶原的变化。结果原代兔髓核细胞在体外传至第5代后,细胞由类软骨细胞样的星形或多角形变为梭形或纤维长条形。番红O染色显示,退变组红色平均染色面积显著低于原代(P<0.05),免疫组织化学染色显示,退变组棕黄色平均染色面积显著低于原代(P<0.05)。结论原代兔髓核细胞在体外传到第5代时由类软骨细胞样变成纤维细胞样,且糖胺聚糖和Ⅱ型胶原的含量显著下降,为进一步构建体外髓核细胞退变模型提供了依据。

Objective To test the different biological characteristics between primary and degenerative New Zealand white rabbit nucleus pulposus cells. Methods Primary rabbit nucleus pulposus cells were obtained and passed on to Generation 5. These cells were divided into 2 groups： primary and degenerative cell group （Geaeration 5 ）. Morphologic change of the cells were observed by an inverted microscope and HE staining. Change of glycosaminoglycan was determined by Safranin O staining. Change of type Ⅱ collagen was determined by immunohistochemistry staining. Results The shape of primary cells looked like star while degenerative cells likespindle by the observation an inverted microscope and HE staining. Mean staining area of safranin O and type Ⅱ collagen of primary cells were significant high than degenerative cells（P 〈 0.05） Conclusisons After primary nucleus polposus cells were passed on to generate 5, morphology of the cells changed from chondrocyte - like to fibrocyte - like. The contents of glycosaminoglyean and type Ⅱ collagen were significantly down too.