山东大学学报（医学版） ›› 2014, Vol. 52 ›› Issue (12): 45-49.doi: 10.6040/j.issn.1671-7554.0.2014.113
石业华, 陈志鹏, 马启奎, 鞠瑞华, 王启荣
SHI Yehua, CHEN Zhipeng, MA Qikui, JU Ruihua, WANG Qirong
摘要： 目的 探索藻酸盐凝胶中高密度培养脂肪间充质干细胞(ADSCs)成软骨细胞诱导后,利用凝胶释放的细胞再培养形成软骨组织的新方法。方法 取大鼠腹股沟脂肪组织,Ⅰ型胶原酶消化得到ADSCs原代培养,传代后采用流式细胞术检测间充质细胞表面CD90、CD44表达鉴定细胞。将第6代细胞高密度培养于藻酸钙凝胶中,用软骨诱导培养基诱导培养2周,将细胞放自凝胶中释放并单层培养成软骨诱导2周,观察细胞变化。培养出的组织块经Ⅱ型胶原酶消化后,行免疫细胞化学染色,检测软骨特异性Ⅱ型胶原蛋白的表达。结果 ADSCs原代呈梭形生长,传代后呈规律排列旋涡状生长,间充质干细胞表面CD90、CD44呈阳性。藻酸钙-ADSCs成软骨诱导2周后,释放出的细胞贴壁后逐渐聚集,形成一定体积的质硬组织块,免疫组化检测显示,新生组织块细胞胞质中软骨特异性Ⅱ型胶原蛋白呈阳性表达,表明大鼠ADSCs向软骨细胞分化。结论 藻酸盐凝胶中高密度培养大鼠ADSCs成软骨细胞诱导后,利用诱导后的细胞再培养可形成新的软骨组织。
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