Angiogenic potential of human mesenchymal stromal cell and circulating mononuclear cell cocultures is reflected in the expression profiles of proangiogenic factors leading to endothelial cell and pericyte differentiation

Abstract

Endothelial progenitors found among the peripheral blood (PB) mononuclear cells (MNCs) are interesting cells for their angiogenic properties. Mesenchymal stromal cells (MSCs) in turn can produce proangiogenic factors as well as differentiate into mural pericytes, making MSCs and MNCs an attractive coculture setup for regenerative medicine. In this study, human bone marrow-derived MSCs and PB-derived MNCs were cocultured in basal or osteoblastic medium without exogenously supplied growth factors to demonstrate endothelial cell, pericyte and osteoblastic differentiation. The expression levels of various proangiogenic factors, as well as endothelial cell, pericyte and osteoblast markers in cocultures were determined by quantitative polymerase chain reaction. Immunocytochemistry for vascular endothelial growth factor receptor-1 and α-smooth muscle actin as well as staining for alkaline phosphatase were performed after 10 and 14 days. Messenger ribonucleic acid expression of endothelial cell markers was highly upregulated in both basal and osteoblastic conditions after 5 days of coculture, indicating an endothelial cell differentiation, which was supported by immunocytochemistry for vascular endothelial growth factor receptor-1. Stromal derived factor-1 and vascular endothelial growth factor were highly expressed in MSC-MNC coculture in basal medium but not in osteoblastic medium. On the contrary, the expression levels of bone morphogenetic protein-2 and angiopoietin-1 were significantly higher in osteoblastic medium. Pericyte markers were highly expressed in both cocultures after 5 days. In conclusion, it was demonstrated endothelial cell and pericyte differentiation in MSC-MNC cocultures both in basal and osteoblastic medium indicating a potential for neovascularization for tissue engineering applications.

Marker Expression of Interstitial Cells in Human Skeletal Muscle: An Immunohistochemical Study

“…29 Vice versa, MSCs have also been reported to be able to develop into pericytes. 30 CD73 and CD105 are among the markers included in the minimal criteria for defining MSC, 31 but in normal muscle, we only found CD73 and CD105 in endothelial cells. Corresponding to our findings, CD73 and CD105 have previously been described as endothelial markers.…”

Donor variability alters differentiation and mechanical cohesion of tissue-engineered constructs with human endothelial/MSC co-culture

“…However, the successful development of both cell types requires specific culture conditions, still discussed nowadays (cell ratio, simultaneous or delayed co-culture, medium composition, etc. 16,[20][21][22]24,37 ), and in particular 3D growth structures appear to be mandatory. 20,24,38 We were thus expecting the calcium phosphate scaffolds used in our system to promote coculture growth and activity despite the absence of specific coating such as collagen, commonly used for primary EC monoculture in vitro.…”

P2 Receptors Influence hMSCs Differentiation towards Endothelial Cell and Smooth Muscle Cell Lineages

“…By profiling the differentiation-relevant P2 expression patterns and stimuli in adipose tissue-derived hMSCs, our data give insight into how to use artificial P2 ligands to improve the quality of re-cellularized scaffolds. There have been several approaches to differentiate hMSCs towards the vascular lineages, e.g., the usage of vectors [40], sphingosine 1-phosphate [41] or the conduction of different coculture models [42][43][44]. All these strategies reached the state of endothelial progenitor cells (EPCs) or used EPCs as cell source.…”