二维和三维培养脐带间充质干细胞的差异基因表达研究  

Differential gene expression of umbilical cord mesenchymal stem cells cultured by two-dimensional and three-dimensional

作　　者：徐哲 戴晓宇 赵莎莎 李树建 李栋[3] XU Zhe;DAI Xiao-yu;ZHAO Sha-sha;LI Shu-jian;LI Dong(Research and Development Department,Jinan Wanquan Biotechnology Co.Ltd,Jinan 250000,China;Research and Development Department,Shandong Quanxi Biotechnology Co.,Ltd.,Jinan 250000,China;Stem Cell and Regenerative Medicine Center of Shandong University,Jinan 250000,China)

机构地区：[1]济南万泉生物技术有限公司研发部,山东250000 [2]山东省泉溪生物技术有限公司研发部,济南250000 [3]山东大学干细胞与再生医学研究中心,济南250000

出　　处：《中国医药生物技术》2024年第4期333-341,共9页Chinese Medicinal Biotechnology

基　　金：山东省科技型中小企业创新能力提升工程项目(2022TSGC2001)。

摘　　要：目的比较二维培养和三维培养的脐带间充质干细胞(UC-MSCs)的细胞形态和基因表达。方法分别采用以聚氟乙烯为支架的三维培养及传统二维培养方法进行稳定增殖期UC-MSCs细胞的培养,使用倒置显微镜、扫描电镜等观察比较二维培养与三维培养在细胞形态、细胞连接等方面的不同;然后应用基因表达谱芯片技术检测二维培养细胞与三维培养细胞的差异表达基因,用KEGG数据库及GO数据库进行生物信息学分析。结果成功分离并培养人UC-MSCs,流式检测高表达CD105、CD73和CD90等。三维培养的UC-MSCs附着在支架的表面生长并向外扩散,支架各层均可见细胞生长,细胞与细胞间连接紧密。基因表达谱芯片分析显示,三维培养的UC-MSCs与二维培养的UC-MSCs相比,升高的基因主要与生物学过程、分子功能及细胞组分相关,且参与了多个细胞信号通路的调控。结论与二维培养相比,三维细胞培养环境更适合脐带间充质干细胞的生长。Objective To compare morphological and gene expression of two-dimensional(2D)and three-dimensional(3D)cultured umbilical cord mesenchymal stem cells(UC-MSCs).Methods The stably proliferating UC-MSCs were cultured using 3D culture with polyvinyl fluoride as a scaffold and traditional 2D culture methods,respectively.The inverted microscope and scanning electron microscope were used to observe and compare the differences in cell morphology and cell connection between 2D and 3D culture.Gene expression microarray technology was then used to detect the differentially expressed genes between 2D and 3D cultured cells,and bioinformatics analysis was performed using the KEGG database and GO database.Results Human UC-MSCs were successfully isolated and cultured,with CD105,CD73 and CD90 being highly expressed as shown by flow cytometry.UC-MSCs cultured in 3D were attached to the surface of the scaffold and grew outwards.Cell growth was observed in all layers of the scaffold,and cell-to-cell connections were tight.Microarray analysis of gene expression profiles showed that,compared with 2D UC-MSCs,the increased genes of 3D UC-MSCs were mainly related to biological processes,molecular functions,and cell components,and were involved in the regulation of multiple cell signaling pathways.Conclusion Compared with 2D culture,the 3D cell culture environment is more suitable for the growth of UC-MSCs.

关 键 词：脐带间充质干细胞 三维细胞培养 二维细胞培养 基因表达谱芯片 

分 类 号：Q813.11[生物学—生物工程]