人脂肪来源干细胞与聚己内酯/壳聚糖支架的生物相容性研究  

Experiment al Study on Human Adipose -Derived Stem Cells Co -cultured with Poly (ε-Caprolactone)/Chitosan Scaffold

作　　者：周哲[1] 吴稼晟[2] 张明[1] 赵阳[1] 周娟[1] 李伟[2] 王忠[1] 孙康[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]上海交通大学材料科学与工程学院金属基复合材料国家重点实验室,上海市200240

出　　处：《组织工程与重建外科杂志》2013年第3期133-136,共4页Journal of Tissue Engineering and Reconstructive Surgery

基　　金：国家自然科学基金项目(81070605);上海交通大学"医工(理)交叉研究基金"(YG2011MS14);上海市教委科研创新项目(09YZ76)

摘　　要：目的观察人脂肪来源干细胞(Human adipose derived stem cells,hADSCs)在聚己内酯/壳聚糖[Poly(ε-capro-lactone)/chitosan,PCL/CS]支架中的生长情况。方法取PCL/CS浸提液培养hADSCs,CCK-8法检测细胞活力,评价支架细胞毒性。hADSCs传代扩增后,接种到PCL/CS支架上,体外培养1周、2周,裸鼠体内培养2周,HE染色观察细胞在支架上的生长情况,免疫组化HLA-Ⅰ监测经裸鼠体内培养后复合支架上细胞的种属来源。结果 hADSCs在PCL/CS浸提液中可保持较高的增殖率,PCL/CS浸提液无细胞毒性。hADSCs终止于PCL/CS支架后,经过体外、内培养,hADSCs均能长入支架的空隙内,且体内培养比体外培养有更多的细胞长入支架。体外培养1周,已有细胞黏附生长在PCL/CS支架的边缘,体外培养2周后,部分细胞渗透到材料内部。体内培养2周后,大量细胞能渗透到材料内部,同时HLA-Ⅰ抗体检测发现,支架材料内部分细胞HLA-Ⅰ阳性表达,说明PCL/CS支架内该部分的细胞来源于hADSCs。结论 PCL/CS支架安全无毒,hADSCs能在PCL/CS支架上较好生长,该支架可用于组织工程膀胱的研究。Objective To observe the growth of human adipose derived stem cells（hADSCs） cultured with Poly（ε caprolactone）/chitosan biomaterials in vitro and in vivo.Methods hADSCs were isolated from human subcutaneous adipose tissue after collagenase digesting,filtrating and centrifuging.The scaffold was prepared by freeze-drying technique and the method of vacuum thermal cross-linking and it’s cytotoxicity was evaluated by CCK-8 reagent tests.Then the hADSCs of passage 3 were seeded onto the PCL/CS scaffolds and the growth of hADSCs cultured in PCL/CS biomaterials was observed by HE staining.The cells in the scaffolds cultured in vivo were detected by immunohistochemistry.Results hADSCs could maintain high increment rate in the leaching solution of the PCL/CS and the PCL/CS scaffolds were non-toxic.The histological study found that hADSCs could grow into the space of the PCL/CS scaffolds after cultured in vitro and in vivo,and there were more cells in the scaffolds cultured in vivo than in vitro.When cultured in vitro,some cells adhered at the edge of the PCL/CS 1 week later and more cells grew into the inside of the scaffolds after 2 weeks.When cultured in vivo,a great deal of cells grew into the scaffolds,and immunohistochemistry results suggested that the cells inside the scaffolds were hADSCs.Conclusion PCL/CS scaffolds are non-toxic and have a good biocompatibility with hADSCs,which can be used as a vehicle for hADSCs in bladder defect reconstruction.

关 键 词：人脂肪来源干细胞 聚己内酯 壳聚糖 支架 组织工程 

分 类 号：Q813.12[生物学—生物工程]