摘要
目的研究人脂肪来源的间充质干细胞（hADSCs）在聚ε-己内酯共聚物上的附着和增殖能力及分化为内皮细胞的能力。方法选择材料孔径为60-80μm（小孔径）和180-200μm（大孔径）,并计算在14 d内的吸水率和降解率。将hADSCs种植在不同孔径的材料上,计算接种率。利用DAPI免疫荧光标记及扫描电镜观察hADSCs的生长及分布情况,用CCK-8法绘制hADSCs增殖曲线。在材料上含有40 ng/mL VEGF和10 ng/mL bFGF的培养基上培养30 d后,用流式细胞术检测vWF和VE-cadherin,免疫荧光检测Flk-1。结果在小及大孔径材料上14 d的吸水率分别为200%和216.7%,降解率为13.3%和21.7%。hADSCs呈长梭形依附于多孔材料表面。小孔径的细胞接种率为98.3%±0.3%,明显大于大孔径的90.3%±1.5%（P〈0.05）;增殖曲线亦是如此。在小孔径上分化50 d后,vWF和VE-cadherin的阳性率分别为80.9%±0.9%和84.3%±1.1%,大多数细胞表面均表达Flk-1。结论 hADSCs较适合在孔径为60~80μm聚ε-己内酯共聚物材料上生长和增殖,并可在材料上有效分化为内皮细胞。
Objective To study on adhesion ability,growth ability and differentiation into endothelial cells of human adipose-derived mesenchymal stem cells（hADSCs） planted on poly-ε-caprolactone material.Methods We chose the poly-ε-caprolactone with pore diameters of 60~80 μm and 180~200 μm.Absorption and degradated rate were calculated at 14th day.After hADSCs implantated in the materials with different diameters,comparison of seeding rate and growth were carried out by immunofluorescence with DAPI and scanning electron microscope（SEM）.CCK-8 method was used to draw the growth curve.Endothelial differentiation with 40 ng/mL VEGF and 10 ng/mL bFGF after 50 days,vWF and VE-cadherin were detected by flow cytometers and Flk-1 was observed by immunofluorescence way.Results Absorption rates of small and large pore-size material at 14th day were respectively calculated as 200% and 216.7%,while degradation rates were 13.3% and 21.7%.After implantation of hADSCs,cells looked long fusiform attaching on the material by SEM.Seeding rate of small pore-size material was 98.3%±0.3%,which was comparatively more than seeding rate as 90.3%±1.5% of large pore-size material,and growth curve showed the same profile.Hence,endothelial differentiation was carried out with small pore-size material.Differentiating after 50 days,positive ration of vWF was 80.9%±0.9% and VE-cadherin was 84.3%±1.1%.Flk-1 expression was shown in most of cells as immuofluorescence.Conclusion Poly-ε-caprolactone material with pore diameter of 60~80 μm is suitable for the growth of hADSCs which successfully differentiated into endothelial cells.
出处
《基础医学与临床》
CSCD
北大核心
2011年第5期490-495,共6页
Basic and Clinical Medicine
基金
科技部社会公益专项(2005DIB1J086)
