Vascular endothelial growth factor transfection induces human bone marrow mesenchymal stem cells differentiating into endothelial-like cells under hypoxia

doi:10.3969/j.issn.2095-4344.2017.09.008

Abstract

Abstract:

BACKGROUND: It has been found that vascular endothelial growth factor can induce the differentiation of bone marrow mesenchymal stem cells into endothelial cells, but can the vascular endothelial growth factor gene promote the differentiation of bone marrow mesenchymal stem cells into vascular endothelial cells in the damaged organ under the hypoxic environment?
OBJECTIVE: To observe whether human bone marrow mesenchymal stem cells induced by vascular endothelial growth factor could differentiate into vascular endothelial cells under hypoxia.
METHODS: The third passage of human bone marrow mesenchymal stem cells were cultured in vitro. Cells in the control group were cultured with conventional culture medium, while those in experimental group were cultured with adenovirus vector containing vascular endothelial growth factor in 5% O2. After 2 weeks of culture, morphological observation and surface-related molecular detection were performed. The levels of vascular endothelial growth factor and endothelial nitric oxide synthase were detected by ELISA. The expression of endothelin and prostacyclin was detected by RT-PCR and western blot assay.
RESULTS AND CONCLUSION: (1) The number of cells in the control group was more than that in the experimental group. The cells in the control group were crowded and arranged irregularly, showing a fiber-like growth, while those in the experimental group were mostly triangular or polygonal, exhibiting a colony-like growth. (2) CD31 was negative in the control group, while CD105 was positive and the positive rate was 99.7%, indicating that the cells still showed the phenotype of bone marrow mesenchymal stem cells. The positive rate of CD31 was significantly increased to 30.33% in the experimental group and the positive rate of CD105 expression was decreased to 58.11%, indicating a typical phenotype of endothelial cells. (3) Compared with the control group, the expression of endothelin, vascular endothelial growth factor and endothelial nitric oxide synthase increased significantly in the experimental group (P < 0.05), and the expression of prostacyclin decreased significantly (P < 0.05). All these findings indicate that vascular endothelial growth factor can promote the differentiation of human bone marrow mesenchymal stem cells into vascular endothelial cells under hypoxia.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Stem Cells;, Mesenchymal Stem Cells, Vascular Endothelial Growth Factors, Endothelial Cells, Tissue Engineering

CLC Number:

R394.2

Hu Ji-hong, Jia Jia, Lu Juan, Wang Qiu-ping, Zhao Jing-miao, Jin Li-mei, Li Jin-juan. Vascular endothelial growth factor transfection induces human bone marrow mesenchymal stem cells differentiating into endothelial-like cells under hypoxia[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(9): 1352-1356.

Figures/Tables 2

References

[1] 祁玥,郑亚宁.血管内皮生长因子的研究进展[J].青海师范大学学报,2010,26(4):61-66.

[2] 刘伟,刘萌,祝劲松,等.人骨髓间充质干细胞的体外培养、鉴定及成骨分化[J].中国组织工程研究,2012,16(14):2515-2519.

[3] 叶兴德,冉茜,邓小军,等.人骨髓间充质干细胞体外培养方法的改良[J].中国输血杂志,2013,26(4):310-315.

[4] 朱雪玲,苏占海.低氧对间充质干细胞生物学功能的影响[J].青海医学院学报,2016,37(1):62-67.

[5] D’Ippolito G,Diabira S,Howard GA,et al.Low oxygen tension inhibits osteogenic differentiation and enhances stemness of human MIAMI Cells.Bone.2006;39(3):513-522.

[6] 柯奇周,于红,杨靖.低氧对骨髓基质细胞向神经细胞分化的影响[J].山西医科大学学报, 2009,6(10):14-16.

[7] 马俊伟.骨髓间充质干细胞向血管内皮细胞诱导分化的实验研究[D].中国医科大学硕士论文,2009.

[8] 孔根现.VEGF分别与bFGF、aFGF联合诱导骨髓间充质干细胞分化为血管内皮细胞[D].南方医科大学硕士论文,2013.

[9] Arutyunyan I,Fatkhudinov T,Kananykhina E,et al.Role of VEGF-A in angiogenesis promoted by umbilical cord-derived mesenchymal stromal/stem cells: in vitro study.Stem Cell Res Ther.2016;7:46.

[10] 王晓娟.丹参多酚酸盐对大鼠骨髓间充质干细胞增殖及血管内皮生长因子表达的影响[J].环球中医药,2013,6(7):492-495.

[11] 曾莉,常以力,邵毅.血管内皮生长因子反义寡脱氧核苷酸联合血管生成素-1对糖尿病视网膜病变大鼠视网膜血管渗漏及新生血管生成的影响[J].眼科新进展,2012,32(10):905-908.

[12] 白志刚,巩凡,马军,等.VEGF165基因转染兔骨髓间充质干细胞治疗兔激素性股骨头坏死的实验研究[J].宁夏医学杂志,2013, 35(10):892-895.

[13] Bandara N,Gurusinghe S,Chen H,et al.Minicircle DNA-mdeiated endothelial nitric oxide synthase gene transfer enhances angiogenic responses of bone marrow-derived mesenchymal stem cells.Stem Cell Res Ther.2016;7:48.

[14] 王智明.内皮源性心血管活性物质与心血管疾病分析[J].中外医学研究,2013,11(6):149-151.

[15] 杨文娟,洪泽,严思敏,等.氧化应激对血管活性物质的影响及其机制研究进展[J].药学研究,2015,34(12):726-730.

[16] 陈芳,王丽,朱中玉,等.内皮素1及其受体在心血管疾病中作用的研究进展[J].中国全科医学,2013,16(9B):3149-3151.

[17] 张珂,陈鹊汀,王晓冬,等.内皮素受体的分布及其生物功能[J].医学研究与教育,2014,31(3):76-81.

[18] 李培武,伏旭,傅仲学.内皮素受体及其受体拮抗剂在心血管疾病中的作用[J].中国现代应用药学,2014,31(3):376-380.

[19] Donckier JE,Michel L,Delos M,et al.Interrelated overexpression of endothelial and inducbile nitric oxide synthases,endothelin-1 and angiogenic factors in papillary thyroid carcinoma.Clin Endocrinol(Oxf).2006;64(6):703-710.

[20] 赵保明,李凤良,宋喜明,等.谷胱甘肽对糖基化终末产物环境中人脐静脉内皮细胞前列环素和内皮素-1分泌的影响[J].郑州大学学报(医学版),2011,46(4):599-601.

[21] Groop PH,Forsblom C,Thomas MC.Mechanisms of disease: Pathway-selective insulin resistance and microvascular complications of diabetes.Nat Clin Pract Endocrinol Metab. 2005;1(2):100.

[22] Ikhapoh IA,Pelham CJ,Agrawal DK,et al.Synergistic effect of angiotensin II on vascular endothelial growth factor-A-mediated differentiation of bone marrow-derived mesenchymal stem cells into endothelial cells. Stem Cell Res Ther. 2015;6:4.