壳聚糖修饰丝素蛋白与人脂肪间充质干细胞体外构建组织工程脂肪  

Construction of tissue engineered adipose using human adipose stem cells with chitosan-modified silk fibroin

作　　者：亢婷[1] 王刚[1] 刘毅[1] 刘刚强[1] 

机构地区：[1]解放军兰州军区兰州总医院全军烧伤整形外科中心,甘肃省兰州市730050

出　　处：《中国组织工程研究》2014年第39期6323-6328,共6页Chinese Journal of Tissue Engineering Research

基　　金：全军医学科研"十二五"重点项目(BWS11C061)~~

摘　　要：背景:在保留丝素蛋白原有优点的基础上,采用带正电荷的水溶性壳聚糖对其表面进行修饰,可改善细胞在支架材料上的黏附性。目的:验证壳聚糖表面修饰丝素蛋白支架材料与人脂肪间充质干细胞的生物相容性及两者体外构建组织工程脂肪的可行性。方法:将第3代人脂肪间充质干细胞悬液以1×107 L-1浓度接种于壳聚糖表面修饰丝素蛋白支架材料上作为实验组,以单纯的细胞悬液为对照组,MTT法检测细胞在支架材料上的黏附和增殖能力。将第3代人脂肪间充质干细胞悬液以1×109 L-1浓度接种于壳聚糖表面修饰丝素蛋白支架材料上,分别进行成脂诱导培养与高糖培养基常规培养,14 d后行细胞-支架复合物油红O染色与RT-PCR检测。结果与结论:人脂肪间充质干细胞在壳聚糖表面修饰丝素蛋白支架材料上黏附、增殖良好。成脂诱导14 d后,油红O染色显示壳聚糖修饰丝素蛋白支架材料上有大量脂肪细胞生成,且过氧化物酶增殖物活化受体γ2基因表达阳性。结果表明壳聚糖表面修饰丝素蛋白支架材料具有良好的体外生物相容性,与人脂肪间充质干细胞共培养可被成功诱导为成熟脂肪细胞。BACKGROUND：Based on the original advantages of silk fibroin, positive charged water-soluble chitosan modified silk fibroin is modified on surface and could improve celladhesion on the scaffolds. OBJECTIVE：To verify the biocompatibility of chitosan-modified silk fibroin with human adipose-derived stem cells （hADSCs）, and feasibility of constructing tissue engineered adipose in vitro. METHODS：The hADSCs at passage 3 were seeded on chitosan-modified silk fibroin at the concentration of 1×107/L, as the experiment group;at the same cellconcentration, hADSCs were seeded in 96-wel plates as the control group. MTT tests were performed to evaluate the adhesion, growth and proliferation of hADSCs on chitosan-modified silk fibroin. Then hADSCs were implanted on the chitosan-modified silk fibroin scaffolds at the concentration of 1×109/L. The hADSCs seeded onto chitosan-modified silk fibroin complexes were respectively cultured with adipogenic differentiation medium and ordinary high-glucose DMEM. The complexes were stained with oil red O, and detected with RT-PCR after cultured 14 days.＆amp;nbsp;RESULTS AND CONCLUSION：The hADSCs adhered to and proliferated on the scaffolds. After cultured with adipogenic differentiation medium for 14 days, oil red O staining demonstrated that there were amount of mature adipocytes on the scaffold. The peroxisome proliferator activated receptorγ2 was positively expressed. The chitosan-modified silk fibroin possessed excellent biocompatibility in vitro. The co-cultured hADSCs could be induced to mature adipocytes successful y.

关 键 词：生物材料 材料相容性 组织工程脂肪 壳聚糖 丝素蛋白 脂肪间充质干细胞 

分 类 号：R318[医药卫生—生物医学工程]