In vitro pro-angiogenic effects of human adipose derived multipotent mesenchymal stromal cells: effect of donor's age

Abstract

Background & Aim Multipotent mesenchymal stromal cells (MSCs) play a pivotal role in regulating blood vessel formation and promoting vascular homeostasis. These effects are primarily mediated through trophic factors secreted by MSCs (MSC secretome). Those include angiogenic, anti-apoptotic, and immunomodulatory factors that are essential for vascular network remodeling. We have previously shown that adipose derived MSCs from elderly subjects have impaired immunomodulatory properties compared to those from young donors. A higher content of proinflammatory factors (e.g., IL-6, IL-8/CXCL8, and MCP-1/CCL2) in the secretome of elderly-MSCs mediates their reduced immunopotency. It is unclear if age-related changes in the MSC secretome impact the pro-angiogenic effects of MSCs. Methods, Results & Conclusion Methods: Human adipose derived MSC at passage 4-5 (P4-P5) from six adult (mean age: 66.5±11.6 years) and six pediatric (mean age: 16.5±3.2) healthy donors were evaluated. The effect of MSCs or MSC conditioned media (MSC-CM) on human umbilical vein endothelial cells (HUVEC, P4-P5) was assessed in: (i) migration assays (i.e. trans-well migration of HUVEC towards MSC-CM) and (ii) tube formation assays (i.e. in vitro Matrigel tube formation assay with MSC-CM and MSC:HUVEC co-cultures). ImageJ and Wimasis image systems were used for analysis. Angiogenic factors (i.e., Angiopoietin-2, EGF, Endoglin, FGF-1, Follistatin, HGF, IL-8, PIGF, VEGF-A, VEGF-C) in the MSC-CM were quantified by multiplex arrays. Results In contrast to pediatric MSC-CM, adult MSC-CM are more effective in inducing HUVEC migration (adult vs pediatric MSC-CM percent increase compared to negative control; mean±SD: 199.1±54.2 vs 80.81±27.6, p Conclusion Adult adipose derived human MSCs perform better than pediatric MSCs in two in vitro angiogenesis assays: HUVEC migration and tube formation. This could relate to age-associated differences in the MSC secretome composition.