Stem cells of adipose rabbit tissue are stimulate into chondrocyte-like phenotype by collagen V in vitro

Abstract

Introduction: Among a variety of biological functions, including an anti-inflammatory effect, collagen V (COLV) regulates the diameter of collagen fibers with an important role in the development of functional tissues. Therefore the aim of this study was to evaluate, in rabbits, the influence of COLV in the induction of differentiation of adipose tissue-derived stem cells to a chondrocyte-like cell phenotype. Materials and Methods: New Zealand Rabbits were used as source of adipose tissues for the isolation of mesenchymal stem cells (MSCs). Preliminary characterization of mesenchymal lineage and differentiation into chondrocyte-like phenotype was confirmed by immunofluorescence analysis using antibodies to collagens I, II (polyclonals), III and CD34 (monoclonals). After 2 and 3 weeks in culture with and without COLV, the cell aggregates were fixed for 2 h in 4% formaldehyde, dehydrated with ethanol, washed with xylene and embedded in paraffin. Different sections were cut and stained with Toluidine blue, Alcian blue and Picrosirius red respectively. Results: Proteoglicans and collagen fibers were observed by assessment of the different stains, confirming the collagen expression. Remarkably, compared to control cultures, in the presence of COLV timulation, MSCs were capable to increase production of collagen I and II, confirming its chondrocyte-like cell phenotype. Conclusion: We conclude that COLV may facilitate the differentiation of rabbit adipose tissue-derived stem cells into a chondrocyte-like phenotype and this result can be considered to be candidate for therapies.