Nonadherent culture method promotes MSC-mediated vascularization in myocardial infarction via miR-519d/VEGFA pathway
Abstract Background Mesenchymal stem cells (MSCs) can provide therapeutic benefits for myocardial infarction (MI) recovery; however, the molecular mechanism by which MSCs improve the heart function is unclear. Methods Microarray analysis was performed to examine the expression profiling of human MSC...
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| Format: | Article |
| Language: | English |
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BMC
2020-07-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13287-020-01780-x |
| _version_ | 1818406317062094848 |
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| author | Baoping Deng Xianlan Zhang Yi Liang Haiming Jiang Weizhao Huang Yinmeng Wu Weiping Deng |
| author_facet | Baoping Deng Xianlan Zhang Yi Liang Haiming Jiang Weizhao Huang Yinmeng Wu Weiping Deng |
| author_sort | Baoping Deng |
| collection | DOAJ |
| description | Abstract Background Mesenchymal stem cells (MSCs) can provide therapeutic benefits for myocardial infarction (MI) recovery; however, the molecular mechanism by which MSCs improve the heart function is unclear. Methods Microarray analysis was performed to examine the expression profiling of human MSCs (hMSCs) grown as adherent cultures (AC-hMSCs) or nonadherent cultures on ultra-low-adherent plates (nonAC-hMSCs). Real-time quantitative polymerase chain reaction (RT-qPCR), western blotting, and enzyme-linked immunosorbent assays (ELISA) were used to assess VEGFA expression and secretion in the AC-hMSCs and nonAC-hMSCs. The paracrine effect of VEGFA-overexpressing AC-MSCs (AC-VEGFA-hMSCs) or VEGFA-knockdown nonAC-hMSCs (nonAC-shVEGFA-hMSCs) on the angiogenic ability of human umbilical vein endothelial cells (HUVECs) was evaluated using tube formation assay. AC-VEGFA-hMSCs or nonAC-shVEGFA-hMSCs were transplanted into myocardial infarction rats to investigate the therapeutic effect of AC-VEGFA-hMSCs or nonAC-shVEGFA-hMSCs. Luciferase reporter assay was used to confirm the association of VEGFA with miR-519d. Results Microarray analysis revealed that VEGFA is downregulated in AC-hMSCs compared to nonAC-hMSCs. Functional assays revealed that high levels of VEGFA produced from AC-VEGFA-hMSCs increased the tube formation capacity of HUVECs in vitro, improved angiogenesis and cardiac performance, and reduced infarct size in a rat MI model. Low levels of VEGFA secretion from nonAC-shVEGFA-hMSCs had the opposite effects. Mechanistically, we found that miR-519d directly targets VEGFA. High levels of VEGFA secreted from VEGFA-overexpressing nonAC-hMSCs abolished the repressive effect of miR-519d on HUVEC angiogenesis. Conclusion Our findings indicate that nonadherent culture-induced secretion of VEGFA plays an important role in MSCs via the miR-519d/VEGFA pathway and may provide a novel therapeutic strategy for MI treatment. |
| first_indexed | 2024-12-14T09:10:01Z |
| format | Article |
| id | doaj.art-cc76f1b94e7940a78d34452439ccf000 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-14T09:10:01Z |
| publishDate | 2020-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-cc76f1b94e7940a78d34452439ccf0002022-12-21T23:08:35ZengBMCStem Cell Research & Therapy1757-65122020-07-0111111410.1186/s13287-020-01780-xNonadherent culture method promotes MSC-mediated vascularization in myocardial infarction via miR-519d/VEGFA pathwayBaoping Deng0Xianlan Zhang1Yi Liang2Haiming Jiang3Weizhao Huang4Yinmeng Wu5Weiping Deng6Department of Cardiothoracic Surgery, Zhongshan People’s HospitalDepartment of Vascular Surgery, Affiliated Hospital of Guilin Medical UniversityDepartment of Cardiothoracic Surgery, Zhongshan People’s HospitalDepartment of Cardiothoracic Surgery, Zhongshan People’s HospitalDepartment of Cardiothoracic Surgery, Zhongshan People’s HospitalDepartment of Cardiothoracic Surgery, Zhongshan People’s HospitalDepartment of Gastroenterology, Taihe Hospital, Hubei University of MedicineAbstract Background Mesenchymal stem cells (MSCs) can provide therapeutic benefits for myocardial infarction (MI) recovery; however, the molecular mechanism by which MSCs improve the heart function is unclear. Methods Microarray analysis was performed to examine the expression profiling of human MSCs (hMSCs) grown as adherent cultures (AC-hMSCs) or nonadherent cultures on ultra-low-adherent plates (nonAC-hMSCs). Real-time quantitative polymerase chain reaction (RT-qPCR), western blotting, and enzyme-linked immunosorbent assays (ELISA) were used to assess VEGFA expression and secretion in the AC-hMSCs and nonAC-hMSCs. The paracrine effect of VEGFA-overexpressing AC-MSCs (AC-VEGFA-hMSCs) or VEGFA-knockdown nonAC-hMSCs (nonAC-shVEGFA-hMSCs) on the angiogenic ability of human umbilical vein endothelial cells (HUVECs) was evaluated using tube formation assay. AC-VEGFA-hMSCs or nonAC-shVEGFA-hMSCs were transplanted into myocardial infarction rats to investigate the therapeutic effect of AC-VEGFA-hMSCs or nonAC-shVEGFA-hMSCs. Luciferase reporter assay was used to confirm the association of VEGFA with miR-519d. Results Microarray analysis revealed that VEGFA is downregulated in AC-hMSCs compared to nonAC-hMSCs. Functional assays revealed that high levels of VEGFA produced from AC-VEGFA-hMSCs increased the tube formation capacity of HUVECs in vitro, improved angiogenesis and cardiac performance, and reduced infarct size in a rat MI model. Low levels of VEGFA secretion from nonAC-shVEGFA-hMSCs had the opposite effects. Mechanistically, we found that miR-519d directly targets VEGFA. High levels of VEGFA secreted from VEGFA-overexpressing nonAC-hMSCs abolished the repressive effect of miR-519d on HUVEC angiogenesis. Conclusion Our findings indicate that nonadherent culture-induced secretion of VEGFA plays an important role in MSCs via the miR-519d/VEGFA pathway and may provide a novel therapeutic strategy for MI treatment.http://link.springer.com/article/10.1186/s13287-020-01780-xMesenchymal stem cellsMyocardial infarctionVEGFAAngiogenesismiR-519dNonadherent culture |
| spellingShingle | Baoping Deng Xianlan Zhang Yi Liang Haiming Jiang Weizhao Huang Yinmeng Wu Weiping Deng Nonadherent culture method promotes MSC-mediated vascularization in myocardial infarction via miR-519d/VEGFA pathway Stem Cell Research & Therapy Mesenchymal stem cells Myocardial infarction VEGFA Angiogenesis miR-519d Nonadherent culture |
| title | Nonadherent culture method promotes MSC-mediated vascularization in myocardial infarction via miR-519d/VEGFA pathway |
| title_full | Nonadherent culture method promotes MSC-mediated vascularization in myocardial infarction via miR-519d/VEGFA pathway |
| title_fullStr | Nonadherent culture method promotes MSC-mediated vascularization in myocardial infarction via miR-519d/VEGFA pathway |
| title_full_unstemmed | Nonadherent culture method promotes MSC-mediated vascularization in myocardial infarction via miR-519d/VEGFA pathway |
| title_short | Nonadherent culture method promotes MSC-mediated vascularization in myocardial infarction via miR-519d/VEGFA pathway |
| title_sort | nonadherent culture method promotes msc mediated vascularization in myocardial infarction via mir 519d vegfa pathway |
| topic | Mesenchymal stem cells Myocardial infarction VEGFA Angiogenesis miR-519d Nonadherent culture |
| url | http://link.springer.com/article/10.1186/s13287-020-01780-x |
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