组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2014, Vol. 10 ›› Issue (6): 309-313.doi: 10.3969/j.issn.1673-0364.2014.06.003

• 论著 • 上一篇    下一篇

人脂肪来源干细胞与膀胱脱细胞基质-丝素蛋白双层支架的生物相容性研究

赵阳，吴稼晟，周哲，周娟，张明，李伟，王忠，孙康，卢慕峻   

  1. 上海交通大学医学院附属第九人民医院泌尿外科；上海交通大学材料科学与工程学院
  • 发布日期:2020-07-23

Human Adipose-De rived Stem Cells and its Biocompatibility with Bladder Acellular Matrix Graft-Silk Fibroin Bilayer Scaffold

ZHAO Yang,WU Jiasheng,ZHOU Zhe,ZHOU Juan,ZHANG Ming,LI Wei,WANG Zhong,SUN Kang,LU Mujun   

  1. Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine;Shanghai Jiaotong University School of Materials Science and Engineering;
  • Published:2020-07-23
  • Contact: 国家自然科学基金项目(81070605，81370860)；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,h ASCs)在膀胱黏膜下脱细胞基质-丝素蛋白(Bladder acellular matrix graft-silk fibroin,BAMG-SF)双层支架材料中的生长情况,分析其生物相容性。方法取h ASCs,置于BAMG-SF浸提液中培养,CCK-8法检测其细胞活力,评价BAMG-SF支架的细胞毒性并绘制生长曲线。扫描电镜观察BAMG-SF双层支架材料的表面形貌。将h ASCs传代扩增后接种到BAMG-SF双层支架材料上,体外培养1周后,转至裸鼠皮下培养1周、2周,HE染色观察细胞在支架上的生长情况。HLA免疫荧光鉴定裸鼠皮下双层支架上细胞的种属来源。结果 h ASCs在BAMG-SF双层支架浸提液中可保持较高的增殖率,根据细胞相对增殖率与细胞毒性分级关系证实BAMG-SF双层支架浸提液无细胞毒性。由h ASCs在BAMG-SF浸提液和DMEM培养基中的生长曲线可知,BAMG-SF有利于h ASCs的生长。将h ASCs接种到BAMG-SF双层支架材料上,经过体外、内培养,h ASCs均能长入支架的空隙内,且体内培养比体外培养有更多的h ASCs细胞长入支架。HLA检测显示支架内细胞部分为h ASCs。结论新型BAMG-SF双层支架材料安全无毒,与h ASCs生物相容性好,可作为细胞载体应用于组织工程膀胱的研究。

关键词: 人脂肪来源干细胞, 膀胱黏膜下脱细胞基质, 丝素蛋白, 双层支架材料, 组织工程

Abstract: Objective To observe the growth of human adipose-derived stem cells （hASCs） in bladder acellular matrix graft-silk fibroin （BAMG-SF） bilayer scaffold and to analyze the biological compatibility of BAMG-SF with hASCs. Methods hASCs were isolated from human subcutaneous adipose tissue after collagenase digesting, filtrating and centrifuging, then cultured in the leaching solution of BAMG-SF. The cytotoxicity of scaffold was evaluated by CCK-8 cell viability assay, and the growth curves were also observed. Surface morphology on BAMG-SF was observed by scanning electron microscopy （SEM）. The hASCs of passage 3 were seeded onto the BAMG-SF bilayer scaffolds for 1 week, then the BAMG-SF bilayer scaffolds seeded with hASCs were transplanted into nude mouse for 1 week or 2 weeks. The growth of cells in BAMG-SF biomaterials was observed by HE staining. The species origin of these cells in the BAMG-SF scaffolds cultured in vivo was detected by Immunofluorescence. Results hASCs maintained high proliferation rate in the leaching solution of BAMG-SF and the BAMG-SF scaffolds were nontoxic absolutely. According to the growth curves of hASCs cultured in the leaching solution of the BAMG-SF and DMEM, BAMG-SF scaffolds were conducive to the growth of hASCs. The histological study found that hASCs could grow into the space of the BAMG-SF scaffolds after cultured in vitro and in vivo. There were more cells in the scaffolds cultured in vivo than in vitro. Immuno-fluorescence suggested that some of the cells inside the scaffolds were hASCs. Conclusion BAMG-SF bilayer scaffolds are nontoxic and have a good biocompatibility with hASCs, which can be used as a vehicle for hASCs in bladder defect reconstruction.

Key words: Human adipose-derived stem cells, Bladder acellular matrix graft, Silk fibroin, Bilayer scaffold, Tissue engineering

中图分类号: