Dermal fibroblasts display similar phenotypic and differentiation capacity to fat-derived mesenchymal stem cells, but differ in anti-inflammatory and angiogenic potential

Abstract

BackgroundMesenchymal stem cells (MSCs) are multipotent stem cells able to differentiate into different cell lineages. However, MSCs represent a subpopulation of a more complex cell composition of stroma cells contained in mesenchymal tissue. Due to a lack of specific markers, it is difficult to distinguish MSCs from other more mature stromal cells such as fibroblasts, which, conversely, are abundant in mesenchymal tissue. In order to find more distinguishing features between MSCs and fibroblasts, we studied the phenotypic and functional features of human adipose-derived MSCs (AD-MSCs) side by side with normal human dermal fibroblasts (HNDFs) in vitroMethodsAD-MSCs and HNDFs were cultured, expanded and phenotypically characterized by flow cytometry (FC). Immunofluorescence was used to investigate cell differentiation. ELISA assay was used to quantify angiogenic factors and chemokines release. Cultures of endothelial cells (ECs) and a monocyte cell line, U937, were used to test angiogenic and anti-inflammatory properties.ResultsCultured AD-MSCs and HNDFs display similar morphological appearance, growth rate, and phenotypic profile. They both expressed typical mesenchymal markers-CD90, CD29, CD44, CD105 and to a minor extent, the adhesion molecules CD54, CD56, CD106 and CD166. They were negative for the stem cell markers CD34, CD146, CD133, CD117. Only aldehyde dehydrogenase (ALDH) was expressed. Neither AD-MSCs nor HNDFs differed in their multi-lineage differentiation capacity; they both differentiated into osteoblast, adipocyte, and also into cardiomyocyte-like cells. In contrast, AD-MSCs, but not HNDFs, displayed strong angiogenic and anti-inflammatory activity. AD-MSCs released significant amounts of VEGF, HGF and Angiopoietins and their conditioned medium (CM) stimulated ECs proliferation and tube formations. In addition, CM-derived AD-MSCs (AD-MSCs-CM) inhibited adhesion molecules expression on U937 and release of RANTES and MCP-1. Finally, after priming with TNFα, AD-MSCs enhanced their anti-inflammatory potential; while HNDFs acquired pro-inflammatory activity.ConclusionsAD-MSCs cannot be distinguished from HNDFs in vitro by evaluating their phenotypic profile or differentiation potential, but only through the analysis of their anti-inflammatory and angiogenic properties. These results underline the importance of evaluating the angiogenic and anti-inflammatory features of MSCs preparation. Their priming with inflammatory cytokines prior to transplantation may improve their efficacy in cell-based therapies for tissue regeneration.

Factors Secreted by Mesenchymal Stem Cells and Endothelial Progenitor Cells Have Complementary Effects on Angiogenesis In Vitro

“…Therefore, it is generally believed that the major benefits derived from transplanted stem cells are the paracrine effects of secreted molecules [17]. This hypothesis was confirmed by numerous studies showing that the CM, collected from a variety of stem cell populations, had beneficial effects on cells and tissues in vitro and in vivo [3,24]. These studies showed that various stem/ progenitor cell populations, such as MSCs or EPCs, released proangiogenic and antiapoptotic cytokines in culture, which may contribute to enhanced angiogenesis and endogenous repair [25].…”

Effects of endothelial cells on human mesenchymal stem cell activity in a three-dimensional in vitro model

“…In a recent publication, the authors identifi ed the diffi culty of distinguishing between adiposederived MSCs (AD-MSCs) and human adult dermal fibroblasts in vitro -regarding their morphological appearance, phenotypic profile and differentiation potential. Both AD-MSCs and HDFas displayed similar capacity to differentiate into osteogenic, adipogenic and cardiomyogenic cell lineages (Blasi et al, 2011). Similarly, dermal-derived fi broblasts from rodent and human were shown to differentiate along the adipogenic and osteogenic lineages (Lorenz et al, 2008).…”

Mesenchymal stem cells in mammary adipose tissue stimulate progression of breast cancer resembling the basal-type

“…32,33 We demonstrate that bMSCs consistently and efficiently chemoattract human endothelial cells in vitro, suggesting their potential involvement in promoting tumor angiogenesis. We also Figure 6.…”