Cells derived from porcine aorta tunica media show mesenchymal stromal-like cell properties in in vitro culture

Abstract

Several studies have already described the presence of specialized niches of precursor cells in vasculature wall, and it has been shown that these populations share several features with mesenchymal stromal cells (MSCs). Considering the relevance of MSCs in the cardiovascular physiopathology and regenerative medicine, and the usefulness of the pig animal model in this field, we reported a new method for MSC-like cell isolation from pig aorta. Filling the vessel with a collagenase solution for 40 min, all endothelial cells were detached and discarded and then collagenase treatment was repeated for 4 h to digest approximately one-third of the tunica media. The ability of our method to select a population of MSC-like cells from tunica media could be ascribed in part to the elimination of contaminant cells from the intimal layer and in part to the overnight culture in the high antibiotic/antimycotic condition and to the starvation step. Aortic-derived cells show an elongated, spindle shape, fibroblast-like morphology, as reported for MSCs, stain positively for CD44, CD56, CD90, and CD105; stain negatively for CD34 and CD45; and express CD73 mRNA. Moreover, these cells show the classical mesenchymal trilineage differentiation potential. Under our in vitro culture conditions, aortic-derived cells share some phenotypical features with pericytes and are able to take part in the formation of network-like structures if cocultured with human umbilical vein endothelial cells. In conclusion, our work reports a simple and highly suitable method for obtaining large numbers of precursor MSC-like cells derived from the porcine aortic wall.

Vascular Regeneration by Stem/Progenitor Cells

“…50 Taken together, the current findings demonstrate a great potential to use resident stem cells for vascular regeneration and angiogenesis. The mechanism of stem/progenitor cell differentiation into endothelial or smooth muscle cells may involve in the alteration of biomechanical stimuli, that is, shear stress and cyclic strain.…”

In vitro differentiation of porcine aortic vascular precursor cells to endothelial and vascular smooth muscle cells

“…The cells were expanded to the further passage when a 60 -65% confluency was reached. Cell doubling and doubling time between passages were calculated as previously described (51).…”

“…Phenotypical characterization of P3 cells was carried out through immunocytochemistry and flow cytometry following the same protocols described by us (51).…”

“…Therefore, in the present work, we decided to modify the previous protocol (51), and we cultured pAVPCs in a pericytespecific medium (pericyte growth medium, PGM), which was successfully used by other authors (3). Indeed, we verified with these new culture conditions MSC-like and pericyte-like properties of pAVPCs.…”

“…We named these cells porcine aortic vascular precursor cells (pAVPCs) for their differentiation potential and for their pericyte-like proprieties in vitro (51).…”