摘要
目的:比较种植于硅橡胶膜或塑料培养板的大鼠纤维环细胞的表型特征.方法:利用倒置相差显微镜及透射电镜观察种植于不同基质的大鼠纤维环细胞形态的变化;利用甲苯胺蓝染色观检测蛋白多糖的表达情况;通过RT-PCR法检测Ⅰ,Ⅱ型胶原及聚集蛋白聚糖mRNA的表达情况;采用流式细胞术观察细胞表面整合素β1的表达情况;同时检测了纤维环细胞在硅橡胶膜上的黏附情况.结果:种植于不同基质的纤维环细胞形态学观察无明显差别;在两种基质上的纤维环细胞甲苯胺蓝染色无差异;Ⅰ,Ⅱ型胶原及聚集蛋白聚糖mRNA及细胞膜整合素β1的表达亦无明显差异;纤维环细胞可以黏附于硅橡胶膜上生长.结论:种植于硅橡胶膜上的纤维环细胞表型特征无改变,为进一步研究力学刺激对纤维环细胞的影响提供实验基础.
AIM： To compare the phenotypic characteristics of rat annulus fibrosus （AF） cells cultured on flexible silicone membranes with the same cells cultured in plastic plates, METHODS： The morphology of AF cells cultured in different substrates was observed under an inverted phase-contrast microscope and a transmission electron microscope. Proteoglycan was stained with toluidine blue for observing its expression. The contents of collagen type Ⅰ , Ⅱ and aggrecan mRNA were determined by RT-PCR. The expression of integrin β1 was monitored by flow cytometry. Cell adhesion to silicone membranes was also measured. RESULTS： The AF cells cultured in different substrates were morphologically undistinguishable. Toluidine blue staining showed that there was also no difference between AF cells cultured on flexible silicone membranes and in plastic plates. The cells growing in different substrates expressed the same levels of collagen type Ⅰ ,Ⅱ and aggrecan mRNA and integrin β1. AF cells grew well on silicone membranes. CONCLUSION： AF cells cultured on flexible silicone membranes maintain the stability of phenotype and may be appropriate for further studying the metabolic responses to mechanical stimuli at the cellular level.
出处
《第四军医大学学报》
北大核心
2006年第13期1169-1172,共4页
Journal of the Fourth Military Medical University
关键词
纤维环细胞
硅橡胶膜
逆转录-聚合酶链式反应
流式细胞术
annulus fibrosus cell
silicone membrane
reverse transcriptase-polymerase chain reaction
flow cytometry
