IntroductionMesenchymal stem cells (MSCs) transplantation has been demonstrated to be an effective strategy for the treatment of cardiovascular disease. However, the low survival rate of MSCs at local diseased tissue reduces the therapeutic efficacy.
We therefore investigated the influence of MicroRNA-378 (miR-378) transfection on MSCs survival and vascularization under hypoxic-ischemic condition in vitro.
Methods:
MSCs were isolated from bone marrow of Sprague-Dawley rats and cultured in vitro. The third passage of MSCs were divided into the miR-378 group and control group.
For the miR-378 group, cells were transfected with miR-378 mimic. Both groups experienced exposure to hypoxia (1% O2) and serum deprivation for 24 hours, using normoxia (20% O2) as a negative control during the process.
After 24 hours of reoxygenation (20% O2), cell proliferation and apoptosis were evaluated. Expressions of apoptosis and angiogenesis related genes were detected.
Both groups were further co-cultured with human umbilical vein endothelial cells to promote vascular differentiation for another 6 hours. Vascular density was assessed thereafter.
Results:
Compared with the control group, MSCs transfected with miR-378 showed more rapid growth.
Their proliferation rates were much higher at 72 h and 96 h under hypoxic condition (257.33% versus 246.67%, P
The miR-378 group formed a larger number of vascular branches on matrigel. BCL2 level was decreased accompanied with an upregulated expression of BAX in the two experimental groups under the hypoxic environment.
BAX expression was reduced in the miR-378 group under the hypoxic environment. In the miR-378 group, there was a decreased expression of tumor necrosis factor-alpha on protein level and a reduction of TUSC-2 under normoxic environment.
Their expressions were both downregulated under hypoxic environment. For the angiogenesis related genes, enhanced expressions of vascular endothelial growth factoralpha, platelet derived growth factor-beta and transforming growth factor-beta1 could be detected both in normoxic and hypoxic-ischemic conditions.
Conclusion:
MiR-378 transfection could effectively promote MSCs survival and vascularization under hypoxic-ischemic condition in vitro.
Author: Yue XingJingying HouTianzhu GuoShaoxin ZhengChangqing ZhouHui HuangYuyang ChenKan SunTingting ZhongJingfeng WangHonghao LiTong Wang
Credits/Source: Stem Cell Research Therapy 2014, 5:130
Published on: 2014-11-23
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