脂肪与华氏果冻间充质干细胞条件培养基对缺氧条件下Versican基因表达影响的比较研究

Q2 Biochemistry, Genetics and Molecular Biology Iranian Biomedical Journal Pub Date : 2022-05-01 DOI:10.52547/ibj.26.3.202
M. Khani, B. Burke, Marzieh Ebrahimi, S. Irani, Fattah Sotoodehnejad
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引用次数: 0

摘要

背景:间充质干细胞通过移植后的旁分泌作用增强组织修复。versican蛋白是促成这种修复机制的重要因素之一。利用间充质干细胞条件培养基培养单核细胞可增加细胞质蛋白的产生,是间充质干细胞移植的良好选择。本研究探讨了人间充质干细胞培养基在模拟缺氧和常氧条件下对骨髓间充质干细胞中versican基因表达的影响。方法:使用的条件培养基来源于脂肪组织或WJ。流式细胞术检测表面标记物(CD105、CD73和CD90)以确认MSCs。分离PBMCs,用来源于脂肪组织或WJ的MSC培养基培养。将去铁胺和氯化钴(终浓度分别为200µM和300µM)添加到单核细胞培养基中诱导模拟缺氧条件。应用Western blotting检测HIF-1α蛋白,确认PBMC的模拟缺氧状态。采用RT-PCR技术检测PBMC中Versican基因的表达。Western blot结果显示,HIF-1α在PBMC中的表达显著升高(p < 0.01)。结果:RT-PCR结果显示,低氧条件下PBMC中versican和VEGF基因的表达较常氧条件下明显升高(p < 0.01)。结论:根据本研究结果,直接利用间充质干细胞替代其分泌因子可改善受损组织。
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Comparison Study on the Effect of Mesenchymal Stem Cells-Conditioned Medium Derived from Adipose and Wharton’s Jelly on Versican Gene Expression in Hypoxia
Background: Mesenchymal stem cells enhance tissue repair through paracrine effects following transplantation. The versican protein is one of the important factors contributing to this repair mechanism. Using MSC conditioned medium for cultivating monocytes may increase versican protein production and could be a good alternative for transplantation of MSCs. This study investigates the effect of culture medium conditioned by human MSCs on the expression of the versican gene in PBMCs under hypoxia-mimetic and normoxic conditions. Methods: The conditioned media used were derived from either adipose tissue or from WJ. Flow cytometry for surface markers (CD105, CD73, and CD90) was used to confirm MSCs. The PBMCs were isolated and cultured with the culture media of the MSC derived from either the adipose tissue or WJ. Desferrioxamine and cobalt chloride (200 and 300 µM final concentrations, respectively) were added to monocytes media to induce hypoxia-mimetic conditions. Western blotting was applied to detect HIF-1α protein and confirm hypoxia-mimetic conditions in PBMC. Versican gene expression was assessed in PBMC using RT-PCR. Western blotting showed that the expression of HIF-1α in PBMC increased significantly (p < 0.01). Results: RT-PCR results demonstrated that the expression of the versican and VEGF genes in PBMC increased significantly (p < 0.01) in hypoxia-mimetic conditions as compared to normoxia. Conclusion: Based on the findings in the present study, the secreted factors of MSCs can be replaced by direct use of MSCs to improve damaged tissues.
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来源期刊
Iranian Biomedical Journal
Iranian Biomedical Journal Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)
CiteScore
3.20
自引率
0.00%
发文量
42
审稿时长
8 weeks
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