Characterization of mesenchymal stem cells derived from equine adipose tissue

A.M Carvalho,L.E.C Álvarez,E Deffune,A.L.G Alves,M.A Golim,A.L.M Yamada,M.L Conceição,L.L Jorge,C.A Hussni

doi:10.1590/s0102-09352013000400001

Abstract

Stem cell therapy has shown promising results in tendinitis and osteoarthritis in equine medicine. The purpose of this work was to characterize the adipose-derived mesenchymal stem cells (AdMSCs) in horses through (1) the assessment of the capacity of progenitor cells to perform adipogenic, osteogenic and chondrogenic differentiation; and (2) flow cytometry analysis using the stemness related markers: CD44, CD90, CD105 and MHC Class II. Five mixed-breed horses, aged 2-4 years-old were used to collect adipose tissue from the base of the tail. After isolation and culture of AdMSCs, immunophenotypic characterization was performed through flow cytometry. There was a high expression of CD44, CD90 and CD105, and no expression of MHC Class II markers. The tri-lineage differentiation was confirmed by specific staining: adipogenic (Oil Red O), osteogenic (Alizarin Red), and chondrogenic (Alcian Blue). The equine AdMSCs are a promising type of adult progenitor cell for tissue engineering in veterinary medicine.

Highlights

The clinical use of cellular therapy and tissue engineering in veterinary medicine is developing principle of adhesion, which allows isolation and characterization of a homogeneous cell population for further use in cell therapy (Pascucci et al, 2011).Traditionally, the process of isolating the nucleated cell fraction from the adipose tissue is based on collagenase digestion, followed by a series of centrifugation steps to isolate the specific cells
The separation of equine adipose-derived mesenchymal stem cells (AdMSCs) from the stromal vascular fraction (SVF) is based on the Several studies reported the characterization of AdMSCs in horses, where the most common processes are osteogenic, adipogenic and chondrogenic differentiation of progenitor cells (Vidal et al, 2007, 2008; Kisiday et al, 2008; Mambelli et al, 2009; Colleoni et al, 2009; Toupadakis et al, 2010; Ahern et al, 2011; Schwarz et al, 2011a,b)
Adipose tissue was successfully collected and a mean of 2g of material was obtained followed by isolation of AdMSCs

Summary

Introduction

The process of isolating the nucleated cell fraction from the adipose tissue is based on collagenase digestion, followed by a series of centrifugation steps to isolate the specific cells. This cell fraction is referred to as stromal vascular fraction (SVF). Five recent studies have reported the analyses of surface antigens of AdMSCs by flow cytometry (de Mattos Carvalho et al, 2009; Braun et al, 2010; Pascucci et al, 2011; Raabe et al, 2011; Ranera et al, 2011). The results from these studies are demonstrated in table 1

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