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

Dermal fibroblasts display similar phenotypic and differentiation capacity to fat-derived mesenchymal stem cells, but differ in anti-inflammatory and angiogenic potential - Download this document for free, or read online. Document in PDF available to download.

Vascular Cell

, 3:5

First Online: 08 February 2011Received: 06 August 2010Accepted: 08 February 2011


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 vitro

MethodsAD-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.

List of abbreviations usedAD-MSCs Adipose-derived MSCs

AD-MSCs-CM conditioned medium-derived AD-MSCs

ALDH aldehyde dehydrogenase

Ang-1 Angiopoietin-1

Ang-2 Angiopoietin-2

AP Alkaline phosphatase

APCs antigen presenting cells

BM bone marrow

BM-MSC s bone marrow derived MSCs

CM conditioned medium

Cx43 Connexin 43

ECs endothelial cells

FCFlow cytometry

HGF Hepatocyte growth factor

HNDFs Human normal dermal fibroblasts

HNDFs-CM conditioned medium-derived HNDF

HMECs human microvascular endothelial cells

HUVECs human umbilical vein endothelial cells

MSCs mesenchymal stem cells

MyoA myocardial actin

LPS lipopolisaccaride

P-AD-MSCs-CM priming-derived AD-MSCs-CM

PDGF platelet derived growth factor

P-HNDFs-CM priming-derived HNDFs-CM

SMCs smooth muscle cells

TGFβ1 transforming growth factor β1

TNFα tumor necrosis factor α

TNP-C cardiac troponin

VEGF vascular endothelial growth factor

vWf von Willebrand factor

Electronic supplementary materialThe online version of this article doi:10.1186-2045-824X-3-5 contains supplementary material, which is available to authorized users.

Antonella Blasi, Carmela Martino contributed equally to this work.

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Author: Antonella Blasi - Carmela Martino - Luigi Balducci - Marilisa Saldarelli - Antonio Soleti - Stefania E Navone - Laura Canz


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