中国组织工程研究

中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (39): 6323-6328.doi: 10.3969/j.issn.2095-4344.2014.39.016

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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

亢  婷，王  刚，刘  毅，刘刚强   

  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
  • 出版日期:2014-09-17 发布日期:2014-09-17
  • 通讯作者: 刘毅，主任医师，教授，硕士生导师，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
  • 作者简介:亢婷，女，1988年生，河南省新乡市人，汉族，在读硕士，主要从事脂肪组织工程研究。
  • 基金资助:

    全军医学科研“十二五”重点项目(BWS11C061)

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

Kang Ting, Wang Gang, Liu Yi, Liu Gang-qiang   

  1. PLA Center for Military Burns and Plastic Surgery, Lanzhou General Hospital, Lanzhou Command of Chinese PLA, Lanzhou 730050, Gansu Province, China
  • Online:2014-09-17 Published:2014-09-17
  • Contact: Liu Yi, Chief physician, Professor, Master’s supervisor, PLA Center for Military Burns and Plastic Surgery, Lanzhou General Hospital, Lanzhou Command of Chinese PLA, Lanzhou 730050, Gansu Province, China
  • About author:Kang Ting, Studying for master’s degree, PLA Center for Military Burns and Plastic Surgery, Lanzhou General Hospital, Lanzhou Command of Chinese PLA, Lanzhou 730050, Gansu Province, China
  • Supported by:

    Key Project of Military Medical Research during the Twelfth Five-Year Plan, No. BWS11C061

PDF

580

被引次数

2

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。


中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程


全文链接：

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

Abstract:

BACKGROUND: Based on the original advantages of silk fibroin, positive charged water-soluble chitosan modified silk fibroin is modified on surface and could improve cell adhesion 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 1×10 7/L, as the experiment group; at the same cell concentration, hADSCs were seeded in 96-well 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 1×10 9/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.
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 receptor γ2 was positively expressed. The chitosan- modified silk fibroin possessed excellent biocompatibility in vitro. The co-cultured hADSCs could be induced to mature adipocytes successfully.


中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程


全文链接：

Key words: chitosan, silk fibroin, mesenchymal stem cells

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