Stabilization of Hypoxia-Inducible Factor-1 alpha enhances the tubulogenic capacity of genetically modified Muse cells

NUÑEZ PEDROZO CN; CUNIBERTI L; GIMÉNEZ CS; OLEA FD; CASTILLO MG; CROTTOGINI A

Rosario

Congreso; Reunion anual SAFIS Rosario 2019; 2019

SAFIS

Angiogenesis is the formation of new blood vessels from existing vasculature and is an integral part of both normal and pathological processes. Muse cells are a subpopulation of Mesenchymal stem cells (ASC) that has been reported to repair acute myocardial infarction (AMI) in animal models. The objective of our study was to improve genetically human Muse cells transduced with the double-mutated HIF-1a gene to assess their angiogenic ability in an in-vitro tube formation assay. Methods. ASC and Muse cells were isolated from human lipoaspirates. In a third group, Muse cells were modified with a baculovirus that encodes a mutant form of the HIF1a transcription factor that does not degrade under normoxia conditions (dmHIF-1a-Muse). Changes in the expression of the angiogenic genes HIF-1a, VEGF and FGF were studied by RT-qPCR. Additionally, in vitro tube formation assay was performed and the rings formed per unit area were quantified. Results. HIF-1a expression increased 5.9 fold in Muse and 2555 fold in Muse-HIF compared to its expression in ASCs. VEGF and FGF expression showed similar behavior; VEGF 3.3 and 7.9 and FGF 3.5 and 5.9 fold increases, respectively. In the tubulogenesis test, significant differences were observed in the number of rings per area (mm2) between dmHIF-1a-Muse group (31.42 ± 12.47) compared to the Muse group (21.75 ± 12.50, p˂0.05) and ASC group (21.33 ± 5.63 p˂0.01), One way ANOVA Bonferroni. However non-significant difference was observed between Muse and ASC groups. Conclusion. Muse cells modified with dmHIF-1a increased their angiogenic capacity at the level of expression of molecules involved in the proliferation of vascular endothelial cells, as well as in the formation of three-dimensional vessels in functional assays