摘要
目的探究腺病毒介导的IL-8受体过表达对脐带源间充质干细胞生物活性的影响。方法本研究利用含有IL-8RA和IL-8RB的c DNA和绿色荧光蛋白(GFP)基因的重组腺病毒载体(p Ad-IL-8RA-GFP、p Ad-IL-8RB-GFP和p Ad-Null-GFP)分别转染分离培养的P3-5代脐带源间充质干细胞,倒置荧光显微镜观察GFP的表达,流式细胞仪检测干细胞标签蛋白CD29、CD44、CD34、HLA-DR的表达,CCK-8实验检测转染后细胞的增殖能力,粘附实验及Transwell趋化实验观察过表达IL-8RA/B的脐带源间充质干细胞的迁移能力。结果脐带源间充质干细胞形态均一、生长状态良好;干细胞特异性相关蛋白CD29、CD44阳性表达,CD34、HLA-DR阴性表达;与对照组比较,IL-8RA/B过表达的脐带源间充质干细胞的增殖能力无明显改变,但是过表达IL-8RA/B的间充质干细胞粘附在损伤内皮上以及迁移到小室下方的细胞数目更多。结论以上结果表明IL-8RA/B的过表达不影响脐带源间充质干细胞的增殖能力,但是能够促进脐带源间充质干细胞向IL-8趋化和向损伤内皮细胞粘附。
In order to explore the effects of the overexpression of IL-8 receptor on the biological activity of mesenchymal stem cells derived from human umbilical cord（h UC-MSCs）. Methods In this study, passage 3-5 h UC-MSCs were collected and transfected with adenovirus vectors that contain human IL-8 RA and/or IL-8 RB cDNAs and the green fluorescent protein（GFP） gene, or Ad Null（empty vector） and GFP（pAd-IL-8 RA-GFP, or pAd-IL-8 RB-GFP and pAd-Null-GFP,respectively. Flow cytometry was used to detect surface antigens of primary cultured mesenchy-mal stem cells,CCK-8 assay examined the proliferation of h UC-MSCs before and after transfection, the expression of GFP observed with the inverted fluorescence microscope. Adhesion assay and Transwell assay were carried out to evaluate IL-8 induced migration ability and adheringcapacity of h UC-MSCs over expressing IL-8 RA/B. R esults showed that h UC-MSCs were uniform in shape, and the expression of CD29 and CD44 were positive, and the expression of CD34 and HLA-DR were negative. The proliferation activity of h UC-MSCs overexpressing IL-8 RA/B wasn＇t changed. Compared with the empty plasmid group,Adhesion assay and transwell assay showed that there was more h UC-MSCs overexpressed IL-8 RA/B that sticked to the injured endothelial cells and migrated to the bottom of the chamber. Conclusion suggest that the overexpression of IL-8 RA/B didn＇t affect the proliferation activity of h UC-MSCs, but significantly enhanced h UC-MSCs＇ migration and adhesion to ECs.
作者
王飒
王文强
李秋实
慕晓玲
Wang Sa;Wang Wenqiang;Li Qiushi;Mu Xiaoling(Key Laboratory of Xinjiang Endemic and Ethnic Diseases/School of Medicine,Shihezi University,Shihezi,Xinjiang 832000,China)
出处
《石河子大学学报（自然科学版）》
CAS
北大核心
2018年第3期293-299,共7页
Journal of Shihezi University(Natural Science)
基金
新疆兵团社会发展科技攻关与成果转化计划(2016AD013)
自治区研究生科研创新项目(XJGRI2016041)
关键词
人脐带源间充质干细胞
IL-8RA/B
腺病毒
细胞迁移或粘附
mesenchymal stem cells derived from human umbilical cord （hUC-MSCs）
IL-8RA/B
adenovirus
migration/adhesion
