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UBM材料上的Laminin蛋白促进负载骨髓间充质干细胞增殖和平滑肌分化

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Keywords: 脱细胞膀胱,骨髓间充质干细胞,Laminin蛋白,细胞增殖,细胞分化?

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Abstract:

目的探讨猪膀胱细胞外基质(urinarybladdermatrix,UBM)上的Laminin蛋白对大鼠骨髓间充质干细胞(bonemarrowmesenchymalstemcells,BMSCs)增殖和向平滑肌分化的影响。方法采用机械分离法、酶消化法及冷冻干燥法等方法对新鲜猪膀胱进行脱细胞处理,采用蛋白与支架共孵育,制备出UBM+Laminin及UBM+Anti-Laminin复合支架。采用贴壁法培养BMSCs,流式细胞仪鉴定BMSCs。采用P3代BMSCs,分别接种于UBM+Laminin、UBM及UBM+Anti-Laminin上。采用CCK-8检测、HE染色观察及免疫组化鉴定等方法检测Laminin蛋白对BMSCs增殖、迁移和分化的影响。结果流式细胞仪鉴定BMSCsCD29阳性表达率为98%,CD31阳性表达率为4.56%,CD34阳性率为3.30%。3种材料均能支持BMSCs生长,且随共孵育时间的延长,D(450)值逐渐升高,其中UBM+Laminin复合材料组细胞D(450)最高(P<0.01)。HE染色结果显示UBM+Laminin组细胞接种第3天形成黏附层,并向深部迁移,第14天可达材料致密层;UBM组第7天仅少许细胞向深层迁移,第14天未达基底层;UBM+Anti-Laminin组仅在支架表层生长。免疫组化检测显示UBM+Laminin组、UBM组、UBM+Anti-Laminin组细胞α-SMA表达分别呈阳性、弱阳性和阴性。结论UBM材料上Laminin蛋白能促进负载BMSCs增殖、迁移、并有诱导其向平滑肌方向分化的作用。?

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