组织工程中去细胞化支架的构建,存在着保留细胞外基质和完全去除供体细胞成分的矛盾,如何评价去细胞化流程的效果是解决的关键。本研究采用以TritonX-100为主,辅助以SDS、酶学法、物理冻融法、高低渗法等方法的新型全肝脏去细胞化流程...组织工程中去细胞化支架的构建,存在着保留细胞外基质和完全去除供体细胞成分的矛盾,如何评价去细胞化流程的效果是解决的关键。本研究采用以TritonX-100为主,辅助以SDS、酶学法、物理冻融法、高低渗法等方法的新型全肝脏去细胞化流程制作去细胞支架,通过免疫组化及电镜观察支架形态结构,ELISA法检测细胞外基质各组分含量,吸光度及Western-blotting法检测供体细胞组分残留量,免疫荧光法检测支架内三维循环培养诱导WBF-344细胞分化。结果表明,该流程很好的保留了胶原蛋白、层粘连蛋白、纤粘连蛋白等细胞外三维结构;氨基聚糖、HGF、VEGF等细胞外基质保存量分别达正常组织的90%及40%;基本去除残留DNA的同时,相比SF.Badylak et al.法,细胞残留组分量如GAPDH、MCH-I及HMGB-1明显下降,仅为正常组织的10%;WBF-344细胞在支架培养系统内生长良好,白蛋白表达逐渐增强。本研究成功构建了新的、较完善的去细胞化支架制作与评价体系,为干细胞分化在肝脏疾病治疗找到一种新的应用模式,展开更多
A simplified method for determining 4 pyrethroid insecticides (Fenpropathrin, Cypermethrin, Fenvalirate, and Deltamethrin) in Chrysanthemi is described. Standards were fortified in to Chrysanthemi (5g) with two levels...A simplified method for determining 4 pyrethroid insecticides (Fenpropathrin, Cypermethrin, Fenvalirate, and Deltamethrin) in Chrysanthemi is described. Standards were fortified in to Chrysanthemi (5g) with two levels, 0.1-1 pp m the pyrethroid pesticides are extracted with petroleum ether and cleaned up by natural aluminum oxide. The extracts are analyzed by gas chromatography equipped with electron capture detector (ECD). Analysis of fortified Chrysanthemi (n = 3) shows recovery range from 99-102 % at 1 pp m level and 93-104 % at 0.1 pp m level. The coefficient of variation of the method at 1 st level was 0.12 and 0.14 for 2 ed level. The detection limits of the method ranged from 0.04pp b to 28 pp b. The method is rapid, sample, and sensitive and is applicable to the determination of Fenpropathrin, Cypermethrin, Fenvalirate, and Deltamethrin in Chrysanthemi.展开更多
文摘组织工程中去细胞化支架的构建,存在着保留细胞外基质和完全去除供体细胞成分的矛盾,如何评价去细胞化流程的效果是解决的关键。本研究采用以TritonX-100为主,辅助以SDS、酶学法、物理冻融法、高低渗法等方法的新型全肝脏去细胞化流程制作去细胞支架,通过免疫组化及电镜观察支架形态结构,ELISA法检测细胞外基质各组分含量,吸光度及Western-blotting法检测供体细胞组分残留量,免疫荧光法检测支架内三维循环培养诱导WBF-344细胞分化。结果表明,该流程很好的保留了胶原蛋白、层粘连蛋白、纤粘连蛋白等细胞外三维结构;氨基聚糖、HGF、VEGF等细胞外基质保存量分别达正常组织的90%及40%;基本去除残留DNA的同时,相比SF.Badylak et al.法,细胞残留组分量如GAPDH、MCH-I及HMGB-1明显下降,仅为正常组织的10%;WBF-344细胞在支架培养系统内生长良好,白蛋白表达逐渐增强。本研究成功构建了新的、较完善的去细胞化支架制作与评价体系,为干细胞分化在肝脏疾病治疗找到一种新的应用模式,
文摘A simplified method for determining 4 pyrethroid insecticides (Fenpropathrin, Cypermethrin, Fenvalirate, and Deltamethrin) in Chrysanthemi is described. Standards were fortified in to Chrysanthemi (5g) with two levels, 0.1-1 pp m the pyrethroid pesticides are extracted with petroleum ether and cleaned up by natural aluminum oxide. The extracts are analyzed by gas chromatography equipped with electron capture detector (ECD). Analysis of fortified Chrysanthemi (n = 3) shows recovery range from 99-102 % at 1 pp m level and 93-104 % at 0.1 pp m level. The coefficient of variation of the method at 1 st level was 0.12 and 0.14 for 2 ed level. The detection limits of the method ranged from 0.04pp b to 28 pp b. The method is rapid, sample, and sensitive and is applicable to the determination of Fenpropathrin, Cypermethrin, Fenvalirate, and Deltamethrin in Chrysanthemi.
