摘要
目的探究通过加入间充质干细胞产生的细胞外基质,从而改变肪来源干细胞的微环境后,使得脂肪来源干细胞增殖转化功能改善的核心分子和作用机制。方法采用生物信息学方法,从基因表达谱芯片数据库中获取12组供体的转录组信息,并分为4组:Ad on NHDF组,记录在聚苯乙烯培养基中加入人新生儿真皮成纤维细胞的细胞外基质后,脂肪来源干细胞的转录组表达情况;Ad on Bm组,记录在聚苯乙烯培养基加入骨髓来源间充质干细胞的细胞外基质后,脂肪来源干细胞的转录组表达情况;Ad on Ad组,记录在聚苯乙烯培养基加入脂肪来源干细胞的细胞外基质后,脂肪来源干细胞的转录组表达情况;Ad on TCPS组,记录脂肪来源干细胞在单纯的聚苯乙烯培养基的转录组表达情况。采用京都基因与基因组百科全书、基因本体、基因集合富集分析等富集分析手段,并利用软件R、Perl、Cytoscape进行差异分析和数据可视化。结果每种间充质干细胞产生其特定的细胞外基质,但在加入不同的细胞外基质培养基培养后的脂肪来源干细胞,都共享部分差异表达的基因和蛋白,如编码胶原家族基因。通过进一步将共享差异基因和功能集团构建蛋白互作关系,最终确定出改变微环境后影响脂肪来源干细胞增殖分化的3个重要分子COL4A1、COL11A1、COL15A1,分子的低表达可能与细胞修复、核苷酸代谢、DNA复制和细胞周期等多种生物学反应和信号通路相关。结论加入间充质干细胞产生的细胞外基质培养基培养的脂肪来源干细胞,编码胶原的基因表达量显著下调,并通过多种途径参与脂肪来源干细胞的增殖分化。
Objective This study aims to explore the potential related gene and mechanism of mesenchymal stem cells (MSCs)-derived extracellular matrix (ECM) promoting the proliferation and differentiation of adipose-derived stem cell (ADSCs), through regulating extracellular microenvironment. Methods Twelve transcriptomes were analysed using Gene Expression Omnibus (GEO) database, and divided into four groups:(1) ADSCs cultured in Tissue Culture Polystyrene (TCPS) medium with normal human dermal fibroblasts (NHDF)-derived ECM,(2) ADSCs cultured in TCPS medium with bone marrow mesenchymal stem cells (BMSCs)-derived ECM.(3) ADSCs cultured in TCPS medium with ADSCs-derived ECM.(4) ADSCs cultured in TCPS medium as the control group. Kyoto Encyclopedia of Genes and Genomes (KEGG), Gene Ontology (GO), and Gene Set Enrichment Analysis (GSEA). Software R, Perl and Cytoscape software were used for differential expression analysis and data visualization. Results Each MSC produced specific extracellular matrix. However, the ADSCs cultured with additive ECM expressed partially different genes and proteins, e. g. The collagen family genes. Furthermore, 3 important molecules: COL4A1, COL11A1, COL15A1 were detected by constructing the interaction relationship between the shared genes and the functional groups. They may affect the proliferation and differentiation of ADSCs by changing microenvironment. The low expression of above molecules, may be related to biological processes and signaling pathways, such as cell repair, nucleotide metabolism, DNA replication, cell cycle, etc. Conclusions The gene expression of collagen encoding were down-regulated, when ADSCs cultured in the medium with additive ECM derived by MSCs. This may significantly affect the proliferation and differentiation of ADSCs through various pathways.
作者
李子超
易晓巍
易成刚
Li Zichao;Yi Xiaowei;Yi Chenggang(Department of Burns and Cutaneous Surgery, First Affiliated Hospital of Air Force Military Medical University, Xijing Hospital , Xi′an 710032 , China;Department of Epidemiology and Health Statistics, West China School of Public Health, Sichuan University, Chengdu 610041, China;Department of Plastic Surgery, First Affiliated Hospital of Air Force Military Medical University , Xijing Hospital , Xi′an 710032 , China)
出处
《中华整形外科杂志》
CAS
CSCD
北大核心
2019年第4期361-366,共6页
Chinese Journal of Plastic Surgery
基金
国家自然科学基金面上项目(81671932)
国家自然科学基金青年项目(81701919).
关键词
脂肪来源干细胞
间充质干细胞
体外培养
编码胶原基因
富集分析
Adipose-derived stem cells
Mesenchymal stem cells
Cell culture
Collagen-encoding gene
Enrichment analysis
