啮齿动物上皮祖细胞与生物材料水凝胶生物相容性的体外研究  

Compatibility of rodent epithelial progenitor cells with biomaterial hydrogels in vitro

作　　者：薛杉[1] 吴钢[2] 罗诗雨[3] 张鹏[4] 张洪钿[4] 法志强[1] 郭燕舞[1] 柯以铨[1] 徐如祥[4] 

机构地区：[1]南方医科大学珠江医院神经外科,神经外科研究所,广州510282 [2]广东省脑功能修复和再生重点实验室南方医科大学珠江医院肿瘤中心 [3]南方医科大学第二临床学院 [4]北京军区总医院附属八一脑科医院,北京100700

出　　处：《中华神经创伤外科电子杂志》2015年第6期26-33,共8页Chinese Journal Of Neurotraumatic Surgery：Electronic Edition

基　　金：国家自然科学基金(u0632008;30801184;30772232;30500526;81501046);广东省自然科学基金(S2011040003565)

摘　　要：目的体外检测培养细胞与生物工程水凝胶材料的相互影响,为体外培养细胞与生物工程水凝胶共移植治疗提供理论依据。方法选择两款不同组分的水凝胶Matrigel和PuraMatrix作为实验材料,以EPCs作为检测细胞,将EPCs培养于不同浓度的Matrigel(10%、20%、30%)或PuraMatrix(0.25%、0.5%、1%)表面,或与不同浓度的Matrigel或PuraMatrix混悬培养,于培养的不同时间观察培养细胞的形态,并用Alamar Blue法检测细胞的增殖情况,以确定材料的细胞毒性。观察EPCs在水凝胶中的成管化状态,以检测细胞是否仍具有生理功能。结果三种浓度Matrigel中30%的Matrigel细胞毒性较大,而三种浓度的PuraMatrix中1%的PuraMatrix细胞毒性较大。EPCs培养于Matrigel表面较混悬其中的细胞增殖率高,而且此差异随Matrigel的浓度增高而增强。EPCs在20%和30%的Matrigel表面培养均能成管化,在10%的Matrigel表面和混悬其中培养7 d,多数细胞沉底。20%的Matrigel中培养7 d能够立体成管化。而30%的Matrigel中,EPCs仅能伸出少数突起,不能成管化。PuraMatrix在0.25%和0.5%的浓度时,EPCs培养于PuraMatrix表面与细胞混悬其中的细胞增殖率相比无明显差异;而在1%时EPCs培养于PuraMatrix表面和混悬其中存活率均很低。EPCs在0.25%和0.5%的PuraMatrix表面培养,细胞能够穿入PuraMatrix中。而在0.25%的PuraMatrix中培养细胞可附着于材料上,但较难成管化,0.5%的PuraMatrix中培养7 d能够成管化。三种细胞密度相比,107个/ml的细胞密度EPCs更容易体外成管化。结论 Matrigel和PuraMatrix在适当的浓度(20%Matrigel和0.5%PuraMatrix)均能与培养细胞良好的相容,并且起到支撑作用,模拟正常生理状态,给细胞提供一个三维生长环境。由于PuraMatrix为人工合成肽产物,降解产物无毒性,可能是更适合移植的生物材料。Objective To test the interaction of in vitro cultured cells and biological engineering hydrogel materials, providing theoretical basis for the treatment strategy of in vitro culturedcells and biological engineering hydrogel transplantation. Methods Two different components hydrogel Matrigel and PuraMatrix were used as experimental material, and EPCs derived as previous method used as detecting cells. At different concentrations of Matrigel （10%, 20%, 30%） or PuraMatrix （1%, 0.5%, 0.25%）, EPCs were cultivated on surface or within the hydrogels. Observe the morphology and detect cell proliferation by Alamar Blue method in different time to determine the cytotoxicity of hydrogel materials. Observe the in vitro angiogenesis ability of EPCs to detect whether cells still have physiological function. Results three concentrations of Matrigel, 30% Matrigel is highest cytotoxicity. Among three PuraMatrix concentration, 1% PuraMatrix is highest cytotoxicity. The cell proliferation rate of EPCs training on surface of Matrigel is high than suspension in Matrigel, and the difference between these two training mode enhanced while the concentration of Matrigel increased. The in vitro angiogenesis ability of EPCs was observed on 20% and 30% Matrigel surface. EPCs descended to the bottom when cultured in or on surface of 10% Matrigel 7 days. The in vitro angiogenesis ability of EPCs was observed in 20% Matrigel, but in 30% Matrigel, EPCs only stretch minority pustute. The cell proliferation rates were no significant differences when EPCs training on surfaee of or in 0.25% and 0.5% PuraMatrix. But when EPCs training on surface of or in 1% PuraMatrix, the survival rate of cell is very low. EPCs can migrat in 0.25% and 0.5% PuraMatrix when cultured on surface. But in 0.25% PuraMatrix, EPCs attached to the material, but had difficult to perform in vitro angiogenesis. The in vitro angiogenesis ability of EPCs was observed in 0.5% PuraMatrix. Among three densitys of EPCs, density of 10^7 cell/ ml in vitro angiogenesis are more

关 键 词：内皮祖细胞 水凝胶 生物相容性 

分 类 号：R318.08[医药卫生—生物医学工程]