title: Dermal fibroblasts display similar phenotypic and differentiation capacity to fat-derived mesenchymal stem cells, but differ in anti-inflammatory and angiogenic potential.
abstract: BACKGROUND
  Mesenchymal 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
  
  
  METHODS
  AD-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.
  
  
  RESULTS
  Cultured 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.
  
  
  CONCLUSIONS
  AD-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.
authors.affiliation: Medestea Research and Production Laboratories, Consorzio Carso, 70010 Valenzano, Bari, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Blasi
authors.affiliation: Medestea Research and Production Laboratories, Consorzio Carso, 70010 Valenzano, Bari, Italy.
authors.forename: Carmela
authors.initials: C
authors.lastname: Martino
authors.affiliation: Medestea Research and Production Laboratories, Consorzio Carso, 70010 Valenzano, Bari, Italy.
authors.forename: Luigi
authors.initials: L
authors.lastname: Balducci
authors.affiliation: Medestea Research and Production Laboratories, Consorzio Carso, 70010 Valenzano, Bari, Italy.
authors.forename: Marilisa
authors.initials: M
authors.lastname: Saldarelli
authors.affiliation: Medestea Research and Production Laboratories, Consorzio Carso, 70010 Valenzano, Bari, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Soleti
authors.affiliation: Cellular Neurobiology Laboratory, Department of Cerebrovascular Diseases, Fondazione IRCCS Neurological Institute "Carlo Besta", 20133 Milan, Italy.
authors.forename: Stefania E
authors.initials: SE
authors.lastname: Navone
authors.affiliation: Cellular Neurobiology Laboratory, Department of Cerebrovascular Diseases, Fondazione IRCCS Neurological Institute "Carlo Besta", 20133 Milan, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Canzi
authors.affiliation: Cellular Neurobiology Laboratory, Department of Cerebrovascular Diseases, Fondazione IRCCS Neurological Institute "Carlo Besta", 20133 Milan, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Cristini
authors.affiliation: Cellular Neurobiology Laboratory, Department of Cerebrovascular Diseases, Fondazione IRCCS Neurological Institute "Carlo Besta", 20133 Milan, Italy.
authors.forename: Gloria
authors.initials: G
authors.lastname: Invernici
authors.affiliation: Cellular Neurobiology Laboratory, Department of Cerebrovascular Diseases, Fondazione IRCCS Neurological Institute "Carlo Besta", 20133 Milan, Italy.
authors.forename: Eugenio A
authors.initials: EA
authors.lastname: Parati
authors.affiliation: Cellular Neurobiology Laboratory, Department of Cerebrovascular Diseases, Fondazione IRCCS Neurological Institute "Carlo Besta", 20133 Milan, Italy.
authors.forename: Giulio
authors.initials: G
authors.lastname: Alessandri