组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2012, Vol. 8 ›› Issue (4): 189-194.doi: 10.3969/j.issn.1673-0364.2012.04.003

• 论著 • 上一篇    下一篇

人脂肪来源干细胞与胶原支架共培养的实验研究

徐明曦，周哲，张明，吴稼晟，李伟，孙康，王忠，赵阳，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金属基复合材料国家重点实验室
  • 发布日期:2020-07-23

The Study of Combination of Human Adipose-Derived Stem Cells with Collagen Scaffold

XU Mingxi1,ZHOU Zhe1,ZHANG Ming1,WU Jiasheng2,LI Wei2,SUN Kang2,WANG Zhong1,ZHAO Yang1,LU Mujun1   

  1. 1 Department of Urology,Shanghai Ninth People’s Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 200011,China;2 State Key Lab of Metal Matrix Composites,Shanghai Jiaotong University School of Materials Science and Engineering,Shanghai 200240,China.
  • Published:2020-07-23
  • Contact: 国家自然科学基金面上项目　　（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： 目的观察评价人脂肪来源干细胞(Human adipose derived stem cells,hADSCs)在胶原支架中的生长情况,为进一步体内组织修复研究提供依据。方法取人抽脂术后脂肪,经胶原酶解、过滤、离心获得hADSCs,代传代扩增后,接种到胶原支架上。细胞-材料复合物分别体外培养1周、2周,裸鼠体内培养2周、4周后,HE染色观察细胞在支架上的生长情况,免疫组化HLA-Ⅰ检测经裸鼠体内培养后的复合物上的细胞的种属来源。结果原代培养的hADSCs呈"梭形"或"成纤维细胞"样,并以克隆团形式生长。免疫荧光Vimentin染色阳性。第3代hADSCs经流式细胞鉴定,CD29、CD44、CD105表达阳性,CD45、CD34表达阴性。胶原支架复合hADSCs经过体外、体内培养,hADSCs均能长入胶原支架的空隙内,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,已有细胞粘附生长在胶原支架的边缘,体外培养2周后,更多的细胞渗透到材料内部。体内培养2周后,大量细胞占据材料的边缘,有部分细胞能渗透到材料内部甚至材料全层。体内培养4周后,大量细胞渗透支架全层。HLA-Ⅰ抗体检测发现,支架材料内部细胞阳性表达,说明胶原支架内部的细胞来源于hADSCs。结论胶原支架与hADSCs具有较好的相容性,可作为hADSCs的载体材料,用于组织工程缺损修复的研究。

关键词: 脂肪来源干细胞, 胶原, 支架, 组织工程

Abstract: Objective To observe the growth of human adipose derived stem cells co-culture with collagen biomaterials in vitro and in vivo.Methods hADSCs were isolated from human subcutaneous adipose tissue after collagenase digesting,filtrating and centrifuging.Then the 3 passage of hADSCs were seeded onto the collagen scaffolds and the growth of hADSCs co-culture with collagen biomaterials were observed by using Haematoxylin-Eosin staining,meanwhile the cells in the scaffolds cultured in vivo were detected by immunohistochemistry.Results Primary cultured hADSCs were spindle-shaped or fibroblasts-liked cells,and it could form the clone groups.Immunofluorescence identification of Vimentin was positive,and Flow cytometry results showed that 3 passage of ADSCs were positive for CD29,CD44,CD105,but negative for CD34,CD45.The histological study found that after cultured both in vitro and in vivo,hADSCs could grow in the space of the collagen scaffolds,moreover there were more cells in the scaffolds cultured in vivo than in vitro.When cultured in vitro,a few cells adhered at the edge of the collage 1 week later and more cells grew into the inside of the scaffolds after 2 weeks.When cultured in vivo,several cells were found at the edge of the scaffolds and parts of cells had grown into the inside even the whole layer of the scaffolds after 2 weeks,at the 4th week,a great deal of cells grew into the whole layer of the scaffolds.Simultaneously,immunohistochemistry results suggested the cells in the scaffolds’ inside cultured in vivo were from hADSCs.Conclusion Collagen scaffolds have a good biocompatibility with hADSCs,which can be used as a vehicle for hADSCs to repair tissue defect.

Key words: Human adipose derived stem cells, Collagen, Scaffold, Tissue engineering

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