Extracellular Vesicles from Human Umbilical Cord-Derived MSCs Affect Vessel Formation In Vitro and Promote VEGFR2-Mediated Cell Survival

Muñiz-García, Ana; Wilm, Bettina; Murray, Patricia

doi:10.3390/cells11233750

Cited by 6 publications

(5 citation statements)

References 55 publications

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“…We have previously established and characterized the mouse mesothelial cells that we derived from mouse omentum primary cultures and transduced with a GFP-lentivirus [24]. The in vitro angiogenesis assay using human dermal fibroblasts and human dermal microvascular endothelial cells had been previously established in our lab [22]. Here, we co-cultured the GFP-MCs with the human cells in the angiogenesis assay to explore the interaction between mesothelial and endothelial cells.…”

Section: Discussionmentioning

confidence: 99%

“…NHDFs (passages [14][15][16][17][18][19] were seeded at 1.5 × 10 4 /well on gelatinized glass-bottomed 24-well plates in FGM and cultured for 3 days [22]. On day 1 of the experiment, 3 × 10 4 /well HDMECs or dT-HDMECs were seeded onto the NHDFs in EGM MV2, either alone or in combination with GFP-MCs, GFP-PCs or GFP-HaCaTs.…”

Section: Basic Angiogenesis Assaymentioning

confidence: 99%

“…In order to analyze the interactions of MCs in angiogenic processes, we turned to a modified in vitro angiogenesis assay of primary human fibroblasts and primary human microvascular endothelial cells [18][19][20][21][22][23] in which stable endothelial tubes form within 4-7 days. We used this system to interrogate the interaction of MCs [24], with the human ECs in the in vitro assay.…”

Section: Introductionmentioning

confidence: 99%

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Mesothelial Cells Exhibit Characteristics of Perivascular Cells in an In Vitro Angiogenesis Assay

Koukorava,

Ward,

Ahmed

et al. 2023

Cells

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Mesothelial cells have been shown to have remarkable plasticity towards mesenchymal cell types during development and in disease situations. Here, we have characterized the potential of mesothelial cells to undergo changes toward perivascular cells using an in vitro angiogenesis assay. We demonstrate that GFP-labeled mesothelial cells (GFP-MCs) aligned closely and specifically with endothelial networks formed when human dermal microvascular endothelial cells (HDMECs) were cultured in the presence of VEGF-A165 on normal human dermal fibroblasts (NHDFs) for a 7-day period. The co-culture with GFP-MCs had a positive effect on branch point formation indicating that the cells supported endothelial tube formation. We interrogated the molecular response of the GFP-MCs to the angiogenic co-culture by qRT-PCR and found that the pericyte marker Ng2 was upregulated when the cells were co-cultured with HDMECs on NHDFs, indicating a change towards a perivascular phenotype. When GFP-MCs were cultured on the NHDF feeder layer, they upregulated the epithelial–mesenchymal transition marker Zeb1 and lost their circularity while increasing their size, indicating a change to a more migratory cell type. We analyzed the pericyte-like behavior of the GFP-MCs in a 3D cardiac microtissue (spheroid) with cardiomyocytes, cardiac fibroblasts and cardiac endothelial cells where the mesothelial cells showed alignment with the endothelial cells. These results indicate that mesothelial cells have the potential to adopt a perivascular phenotype and associate with endothelial cells to potentially support angiogenesis.

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Section: Discussionmentioning

confidence: 99%

Section: Basic Angiogenesis Assaymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Mesothelial Cells Exhibit Characteristics of Perivascular Cells in an In Vitro Angiogenesis Assay

Koukorava,

Ward,

Ahmed

et al. 2023

Cells

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“…Exosomes (Exos), small secretory membrane vesicles secreted by cells, play an important role in intercellular communication patterns by transferring molecules (such as mRNAs, miRNAs, and proteins) from one cell to another in order to regulate physiological and pathological processes 12 . The positive benefits of stem cell transplantation may be mostly due to paracrine processes 13 , 14 . Exos are an important component of mesenchymal stem cell paracrine secretion 15 .…”

Section: Introductionmentioning

confidence: 99%

Enhancement of endothelial function and attenuation of portal vein injury using mesenchymal stem cells carrying miRNA-25-3p

Nie,

Zhang,

Luo

et al. 2024

Sci Rep

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The aims of this study were to determine whether human umbilical cord mesenchymal stem cells (hucMSCs) modified by miRNA-25-3p (miR-25-3p) overexpression could promote venous endothelial cell proliferation and attenuate portal endothelial cell injury. HucMSCs and human umbilical vein endothelial cells (HUVEC) were isolated and cultured from human umbilical cord and characterized. Lentiviral vectors expressing miRNA-25-3p were transfected into hucMSCs and confirmed by PCR. We verified the effect of miR-25-3p-modified hucMSCs on HUVEC by cell co-culture and cell supernatant experiments. Subsequently, exosomes of miR-25-3p-modified hucMSCs were isolated from cell culture supernatants and characterized by WB, NTA and TEM. We verified the effects of miR-25-3p-modified exosomes derived from hucMSCs on HUVEC proliferation, migration, and angiogenesis by in vitro cellular function experiments. Meanwhile, we further examined the downstream target genes and signaling pathways potentially affected by miR-25-3p-modified hucMSC-derived exosomes in HUVEC. Finally, we established a rat portal vein venous thrombosis model by injecting CM-DiR-labeled hucMSCs intravenously into rats and examining the homing of cells in the portal vein by fluorescence microscopy. Histological and immunohistochemical experiments were used to examine the effects of miRNA-25-3p-modified hucMSCs on the proliferation and damage of portal vein endothelial cells. Primary hucMSCs and HUVECs were successfully isolated, cultured and characterized. Primary hucMSCs were modified with a lentiviral vector carrying miR-25-3p at MOI 80. Co-culture and cell supernatant intervention experiments showed that overexpression of miRNA-25-3p in hucMSCs enhanced HUVEC proliferation, migration and tube formation in vitro. We successfully isolated and characterized exosomes of miR-25-3p-modified hucMSCs, and exosome intervention experiments demonstrated that miR-25-3p-modified exosomes derived from hucMSCs similarly enhanced the proliferation, migration, and angiogenesis of HUVECs. Subsequent PCR and WB analyses indicated PTEN/KLF4/AKT/ERK1/2 as potential pathways of action. Analysis in a rat portal vein thrombosis model showed that miR-25-3p-modified hucMSCs could homing to damaged portal veins. Subsequent histological and immunohistochemical examinations demonstrated that intervention with miR-25-3p overexpression-modified hucMSCs significantly reduced damage and attenuated thrombosis in rat portal veins. The above findings indicate suggest that hucMSCs based on miR-25-3p modification may be a promising therapeutic approach for use in venous thrombotic diseases.

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“…Angiogenesis is defined as the growth of new capillaries on pre-existing blood vessels [2]. The regulation of angiogenesis has been extensively studied for the treatment of ischaemic diseases, wound healing, and tissue regeneration [3]. Studies suggest that muscle regeneration defects may be attributed to damaged blood vessels [4].…”

Section: Introductionmentioning

confidence: 99%

A Novel tRNA-Derived Fragment, tRFGlnCTG, Regulates Angiogenesis by Targeting Antxr1 mRNA

Chen,

Shen,

Liao

et al. 2023

IJMS

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As a novel non-coding RNA with important functions corresponding to various cellular stresses, the function of tRFs in angiogenesis remains unclear. Firstly, small RNA sequencing was performed on normal and post-muscle injury mouse tibialis anterior muscle to identify and analyse differentially expressed tRF/tiRNA. tRNA GlnCTG-derived fragments (tRFGlnCTG) were found to be overexpressed in high abundance in the damaged muscle. Subsequent in vitro experiments revealed that the overexpression of tRFGlnCTG suppressed the vascular endothelial cells’ viability, cell cycle G1/S transition, proliferation, migration, and tube-formation capacity. Similarly, in vivo experiments showed that the tRFGlnCTG decreased the relative mRNA levels of vascular endothelial cell markers and pro-angiogenic factors and reduced the proportion of CD31-positive cells. Finally, luciferase activity analysis confirmed that the tRFGlnCTG directly targeted the 3′UTR of Antxr1, leading to a significant reduction in the mRNA expression of the target gene. These results suggest that tRFGlnCTG is a key regulator of vascular endothelial cell function. The results provide a new idea for further exploration of the molecular mechanisms that regulate angiogenesis.

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Extracellular Vesicles from Human Umbilical Cord-Derived MSCs Affect Vessel Formation In Vitro and Promote VEGFR2-Mediated Cell Survival

Cited by 6 publications

References 55 publications

Mesothelial Cells Exhibit Characteristics of Perivascular Cells in an In Vitro Angiogenesis Assay

Mesothelial Cells Exhibit Characteristics of Perivascular Cells in an In Vitro Angiogenesis Assay

Enhancement of endothelial function and attenuation of portal vein injury using mesenchymal stem cells carrying miRNA-25-3p

A Novel tRNA-Derived Fragment, tRFGlnCTG, Regulates Angiogenesis by Targeting Antxr1 mRNA

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