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

• Published in: Cells (Basel, Switzerland) Vol. 12; no. 20; p. 2436
• Main Authors: Koukorava, Chrysa, Ward, Kelly, Ahmed, Katie, Almaghrabi, Shrouq, Dauleh, Sumaya, Pereira, Sofia M, Taylor, Arthur, Haddrick, Malcolm, Cross, Michael J, Wilm, Bettina
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.10.2023; MDPI
• Subjects: Angiogenesis; Biochemical assays; Blood vessels; Cardiomyocytes; Cell culture; Cell growth; cell migration; cell shape; Coculture Techniques; Endothelial cells; Endothelial Cells - metabolism; Endothelium; Epithelial Cells; Extracellular matrix; Fibroblasts; Flow cytometry; Gene expression; Health aspects; Heart; Homeostasis; Humans; in vitro angiogenesis assay; Mesenchymal Stem Cells; mesothelial cells; mesothelial-mesenchymal transition; Mesothelium; Methods; Microvasculature; Neovascularization; Penicillin; Pericytes; perivascular cells; Permeability; Phenotypes; Smooth muscle; Vascular endothelial growth factor; United Kingdom; United Kingdom > UK; United States > US; Germany; Zeb1; cardiac spheroids; mesothelial-mesenchymal transition; cell shape; Ng2; cell migration; mesothelial cells; in vitro angiogenesis assay; perivascular cells
• ISSN: 2073-4409; 2073-4409
• DOI: 10.3390/cells12202436

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-A 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 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 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.