人脂肪来源干细胞与左旋聚乳酸/聚己内酯支架的体内外生物相容性研究  

Biocompatibility on Human Adipose-Derived Stem Cells Co-cultured with Poly-L-lactide/Polycaprolactone Scaffold

作　　者：姚海军[1] 赵阳[1] 周哲[1] 肖冬冬[1] 周娟[1] 张明[1] 王忠[1] 何创龙[2] 卢慕峻[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院泌尿外科,上海市200011 [2]东华大学化学化工与生物工程学院,上海市200051

出　　处：《组织工程与重建外科杂志》2014年第5期255-258,共4页Journal of Tissue Engineering and Reconstructive Surgery

基　　金：国家自然科学基金面上项目(81370860;81070605);上海交通大学"医工(理)交叉研究基金"(YG2011MS14)

摘　　要：目的观察人脂肪来源干细胞(Human adipose-derived stem cells,hADSCs)在左旋聚乳酸/聚己内酯(Poly-Llactide/Polycaprolactone,PLLA/PCL)复合支架材料中的生长情况及其生物相容性。方法体外分离、培养和扩增hADSCs,制备PLLA/PCL复合支架。取PLLA/PCL浸提液培养hADSCs,CCK-8法检测细胞活力,评价支架的细胞毒性。hADSCs传代扩增后,接种到PLLA/PCL支架材料上。体外培养1周,裸鼠皮下分别培养1周、2周,HE染色观察细胞在支架上的生长情况。免疫荧光检测裸鼠体内复合支架材料内细胞平滑肌肌动蛋白(Smooth muscle actin,SMA)表达情况。结果hADSCs在PLLA/PCL浸提液中可保持较高的增殖率,表明PLLA/PCL浸提液无细胞毒性。hADSCs接种于PLLA/PCL支架上,经体外、体内培养后,均能长入PLLA/PCL支架的孔隙内,且体内培养比体外培养有更多的细胞长入支架内部。经体内培养后,复合细胞的支架比单纯支架有更多的细胞渗入支架内部。细胞材料复合物体内培养2周后,免疫荧光检测发现,支架材料内部分细胞SMA表达阳性。结论 PLLA/PCL复合支架材料,安全无毒,与hADSCs生物相容性好,可作为种子细胞的载体用于组织工程膀胱缺损修复的研究。Objective To observe the growth of human adipose-derived stem cells （hADSCs） on Poly-L-lactide/ Polycaprolactone composite scaffold materials and the biological compatibility. Methods hASCs were isolated from human subcutaneous adipose tissue and PLLA/PCL scaffold was prepared. The cytotoxicity of scaffold was evaluated by CCK-8 cell viability assay. The P3 hASCs were seeded onto the PLLMPCL scaffolds. The growth of cell in PLLA/PCL biomaterials in vivo （1 w and 2 w） and in vitro （1 w） was observed by HE staining. The expression of SMA of the seeding cells in vivo was detected by immunofluorescence. Results hADSCs maintained high proliferation rate in the leaching solution of the PLLA/ PCL and the PLLA/PCL scaffolds were nontoxic absolutely. The histological study found that hADSCs could grow into the space of the PLLA/PCL scaffolds after cultured in vitro and in vivo. There were more cells in the scaffolds cuhured in vivo than in vitro, also more cells in the cell-seeded scaffold than simple scaffold. Some of the seeding cells were positive for SMA after 2 weeks implanted in the scaffold in vivo. Conclusion PLLA-PCL composite scaffolds are nontoxic and have a good biocompatibility with hADSCs, which can be used as a vehicle for hADSCs in the repair of bladder defect.

关 键 词：人脂肪来源干细胞 左旋聚乳酸 聚己内酯 生物相容性 

分 类 号：Q813.11[生物学—生物工程]