猪小肠黏膜下层细胞外基质促进肝细胞活力和功能基因表达的研究
发布时间：2018-10-07 21:24
【摘要】：目的探讨猪小肠黏膜下层细胞外基质(porcine small intestinal submucosa extracellular matrix,PSISM)对肝细胞活力及相关基因表达的调控,为PSISM应用于肝脏组织工程提供实验依据。方法实验分为两部分。(1)取PSISM添加至含10%FBS的H-DMEM培养基中,制备终浓度为50、100、200μg/m L的PSISM培养基;将大鼠肝细胞系BRL细胞分别用以上浓度PSISM培养基培养(A1、B1、C1组),以单纯H-DMEM培养基培养作为对照(D1组)。(2)取PSISM溶解于含0.1 mol/L Na OH的PBS无菌溶液中,调整溶液浓度为1%、2%、3%,制备PSISM凝胶;将BRL细胞分别接种于以上浓度PSISM凝胶进行培养(A2、B2、C2组),鼠尾Ⅰ型胶原凝胶作为对照(D2组)。培养后第1、3、5天,采用Live/Dead荧光染色观察细胞形态及存活情况,细胞增殖-毒性检测试剂盒(cell counting kit-8,CCK-8)检测细胞活力,实时荧光定量PCR检测肝细胞特异基因白蛋白(albumin,ALB)、细胞角蛋白18(cytokeratin 18,CK18)和甲胎蛋白(alpha-fetoprotein,AFP)的表达。结果 Live/Dead荧光染色显示各组细胞均存活良好。细胞活力检测显示,PSISM培养基培养后,第5天C1组吸光度(A)值显著高于D1组(P0.05),其余各时间点各组间比较差异均无统计学意义(P0.05);PSISM凝胶培养后,A2、B2、C2组第3天和第5天A值显著高于D2组(P0.05),A2组第5天A值显著高于B2、C2组(P0.05),其余各时间点各组间比较差异均无统计学意义(P0.05)。实时荧光定量PCR检测显示,A1、B1、C1组ALB、CK18基因相对表达量较D1组显著增高(P0.05)、AFP基因相对表达量降低(P0.05),C1组CK18基因相对表达量显著低于A1、B1组(P0.05);A2、B2、C2组ALB、CK18基因相对表达量较D2组显著增高(P0.05)、AFP基因相对表达量降低(P0.05),A2组CK18基因相对表达量显著高于B2组(P0.05)、AFP基因相对表达量显著低于C2组(P0.05)。其余各组间各基因相对量表达比较,差异均无统计学意义(P0.05)。结论 PSISM无毒性,对肝细胞有良好相容性,能促进肝细胞活力和功能基因表达,有望作为细胞培养添加物或细胞移植载体用于肝脏组织工程。
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M%E6%97%A0%E6%AF%92%E6%80%A7%2C%E5%AF%B9%E8%82%9D%E7%BB%86%E8%83%9E%E6%9C%89%E8%89%AF%E5%A5%BD%E7%9B%B8%E5%AE%B9%E6%80%A7%2C%E8%83%BD%E4%BF%83%E8%BF%9B%E8%82%9D%E7%BB%86%E8%83%9E%E6%B4%BB%E5%8A%9B%E5%92%8C%E5%8A%9F%E8%83%BD%E5%9F%BA%E5%9B%A0%E8%A1%A8%E8%BE%BE%2C%E6%9C%89%E6%9C%9B%E4%BD%9C%E4%B8%BA%E7%BB%86%E8%83%9E%E5%9F%B9%E5%85%BB%E6%B7%BB%E5%8A%A0%E7%89%A9%E6%88%96%E7%BB%86%E8%83%9E%E7%A7%BB%E6%A4%8D%E8%BD%BD%E4%BD%93%E7%94%A8%E4%BA%8E%E8%82%9D%E8%84%8F%E7%BB%84%E7%BB%87%E5%B7%A5%E7%A8%8B.
【作者单位】： 四川大学华西医院再生医学研究中心干细胞与组织工程研究室;
【基金】：四川省科技支撑计划资助项目(2013SZ0088)~~
【分类号】：R657.3
本文编号：2255723
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M%E6%97%A0%E6%AF%92%E6%80%A7%2C%E5%AF%B9%E8%82%9D%E7%BB%86%E8%83%9E%E6%9C%89%E8%89%AF%E5%A5%BD%E7%9B%B8%E5%AE%B9%E6%80%A7%2C%E8%83%BD%E4%BF%83%E8%BF%9B%E8%82%9D%E7%BB%86%E8%83%9E%E6%B4%BB%E5%8A%9B%E5%92%8C%E5%8A%9F%E8%83%BD%E5%9F%BA%E5%9B%A0%E8%A1%A8%E8%BE%BE%2C%E6%9C%89%E6%9C%9B%E4%BD%9C%E4%B8%BA%E7%BB%86%E8%83%9E%E5%9F%B9%E5%85%BB%E6%B7%BB%E5%8A%A0%E7%89%A9%E6%88%96%E7%BB%86%E8%83%9E%E7%A7%BB%E6%A4%8D%E8%BD%BD%E4%BD%93%E7%94%A8%E4%BA%8E%E8%82%9D%E8%84%8F%E7%BB%84%E7%BB%87%E5%B7%A5%E7%A8%8B.
【作者单位】： 四川大学华西医院再生医学研究中心干细胞与组织工程研究室;
【基金】：四川省科技支撑计划资助项目(2013SZ0088)~~
【分类号】：R657.3
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