间充质干细胞利用重组血管内皮生长因子-A分化为内皮细胞。
Mesenchymal Stem Cells Differentiate to Endothelial Cells Using Recombinant Vascular Endothelial Growth Factor -A.
作者信息
Khaki Mohsen, Salmanian Ali Hatef, Abtahi Hamid, Ganji Ali, Mosayebi Ghasem
机构信息
Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran.
Department of Microbiology and Immunology, School of Medicine, Arak University of Medical Sciences, Arak, Iran.
出版信息
Rep Biochem Mol Biol. 2018 Apr;6(2):144-150.
BACKGROUND
Vascular endothelial growth factor-A (VEGF-A), an endothelial cell-specific mitogen produced by various cell types, plays important roles in cell differentiation and proliferation. In this study we investigated the effect of recombinant VEGF-A on differentiation of mesenchymal stem cells (MSCs) to endothelial cells (ECs).
METHODS
VEGF-A was expressed in BL21 (DE3) and BL21 pLysS competent cells with the pET32a expression vector. Recombinant VEGF-A protein expression was verified by SDS-PAGE and western blotting. Mesenchymal stem cell differentiation to ECs in the presence of VEGF-A was evaluated by flow cytometry and fluorescence microscopy.
RESULTS
Recombinant VEGF-A was produced in BL21 (DE3) cells at 0.8 mg/mL concentration. Expression of CD31 and CD 144 was significantly greater, while expression of CD90, CD73, and CD44 was significantly less, in MSCs treated with our recombinant VEGF-A than in those treated with the commercial protein (p < 0.05).
CONCLUSION
Recombinant VEGF-A expressed in a prokaryotic system can induce MSCs differentiation to ECs and can be used in research and likely therapeutic applications.
背景
血管内皮生长因子 -A（VEGF -A）是一种由多种细胞类型产生的内皮细胞特异性促有丝分裂原，在细胞分化和增殖中起重要作用。在本研究中，我们调查了重组VEGF -A对间充质干细胞（MSCs）向内皮细胞（ECs）分化的影响。
方法
用pET32a表达载体在BL21（DE3）和BL21 pLysS感受态细胞中表达VEGF -A。通过SDS -PAGE和蛋白质印迹法验证重组VEGF -A蛋白的表达。通过流式细胞术和荧光显微镜评估在VEGF -A存在下间充质干细胞向内皮细胞的分化。
结果
在BL21（DE3）细胞中产生的重组VEGF -A浓度为0.8 mg/mL。与用商业蛋白处理的间充质干细胞相比，用我们的重组VEGF -A处理的间充质干细胞中CD31和CD144的表达显著更高，而CD90、CD73和CD44的表达显著更低（p <0.05）。
结论
在原核系统中表达的重组VEGF -A可诱导间充质干细胞向内皮细胞分化，可用于研究及可能的治疗应用。
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