Phenotypic and functional characterisation of ovine mesenchymal stem cells: application to a cartilage defect model.
BACKGROUND
Multipotent mesenchymal stromal cells (MSC) are of particular interest for their potential clinical use in cartilage engineering, but a consistent model is missing in large animals.


OBJECTIVE
In the absence of any detailed study reporting a complete characterisation of the mesenchymal cells isolated from sheep bone marrow, we fully characterised adherent stromal cells and developed a pre-clinical model of cartilage engineering by implantation of autologous MSC in the Merinos sheep.


METHODS
Ovine MSC (oMSC) were isolated from bone marrow, expanded and further characterised according to the recently proposed definition of the MSC. The experimental model consists of partial-thickness lesions created in the inner part of the patellae of the posterior legs. Lesions were filled with oMSC with or without chitosan, with or without transforming growth factor (TGF)beta-3, in a fibrin clot.


RESULTS
oMSC were shown to display the three main characteristics of MSC: adherence to plastic, phenotypic profile (positive for CD44, CD105, vimentin and negative for CD34 and CD45), and trilineage differentiation potential. We also report two other important functional characteristics of MSC: support of long-term haematopoiesis and immunosuppressive capacity. In vivo, 2 months after implantation the histological analysis revealed chondrocyte-like cells surrounded by a hyaline-like cartilaginous matrix that was integrated to the host cartilage when oMSC were combined with chitosan and TGFbeta-3.


CONCLUSIONS
This study provides for the first time a strong characterisation of oMSC and establishes the basis for a model of cartilage engineering in a large animal.
Mrugala
D
D
Inserm U 844, Montpellier, France.
Bony
C
C
Neves
N
N
Caillot
L
L
Fabre
S
S
Moukoko
D
D
Jorgensen
C
C
Noël
D
D
D000818:Animals; D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D048271:Chitosan; D004195:Disease Models, Animal; D005260:Female; D006410:Hematopoiesis / Q000502:physiology; D007108:Immune Tolerance; D016130:Immunophenotyping; D007718:Knee Injuries / Q000473:pathology / Q000628:therapy*; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000276:immunology* / Q000502:physiology; D010329:Patella / Q000293:injuries*; D034561:Sheep, Domestic; D023822:Tissue Engineering / Q000379:methods; D016896:Treatment Outcome; D014945:Wound Healing
D048271:Chitosan