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Open Access Highly Accessed Research article

Isolation and characterization of ovine mesenchymal stem cells derived from peripheral blood

Jaber Lyahyai12, Diego R Mediano1, Beatriz Ranera1, Arianne Sanz1, Ana Rosa Remacha1, Rosa Bolea3, Pilar Zaragoza1, Clementina Rodellar14 and Inmaculada Martín-Burriel14*

  • * Corresponding author: Inmaculada Martín-Burriel minma@unizar.es

  • † Equal contributors

Author Affiliations

1 Laboratorio de Genética Bioquímica (LAGENBIO), Facultad de Veterinaria, Universidad de Zaragoza, Miguel Servet 177, Zaragoza, 50013, Spain

2 Centre de Génomique Humaine, Faculté de Médecine et Pharmacie, Université Mohammed V Souissi, Rabat, Morocco

3 Centro de Investigación en Encefalopatías y Enfermedades Transmisibles Emergentes (CIEETE), Facultad de Veterinaria, Universidad de Zaragoza, Zaragoza, Spain

4 Instituto Aragonés de Ciencias de la Salud, Zaragoza, Spain

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BMC Veterinary Research 2012, 8:169  doi:10.1186/1746-6148-8-169

Published: 22 September 2012

Abstract

Background

Mesenchymal stem cells (MSCs) are multipotent stem cells with capacity to differentiate into several mesenchymal lineages. This quality makes MSCs good candidates for use in cell therapy. MSCs can be isolated from a variety of tissues including bone marrow and adipose tissue, which are the most common sources of these cells. However, MSCs can also be isolated from peripheral blood. Sheep has been proposed as an ideal model for biomedical studies including those of orthopaedics and transmissible spongiform encephalopathies (TSEs). The aim of this work was to advance these studies by investigating the possibility of MSC isolation from ovine peripheral blood (oPB-MSCs) and by subsequently characterizing there in vitro properties.

Results

Plastic-adherent fibroblast-like cells were obtained from the mononuclear fraction of blood samples. These cells were analysed for their proliferative and differentiation potential into adipocytes, osteoblasts and chondrocytes, as well as for the gene expression of cell surface markers. The isolated cells expressed transcripts for markers CD29, CD73 and CD90, but failed to express the haematopoietic marker CD45 and expressed only low levels of CD105. The expression of CD34 was variable. The differentiation potential of this cell population was evaluated using specific differentiation media. Although the ability of the cultures derived from different animals to differentiate into adipocytes, osteoblasts and chondrocytes was heterogeneous, we confirmed this feature using specific staining and analysing the gene expression of differentiation markers. Finally, we tested the ability of oPB-MSCs to transdifferentiate into neuronal-like cells. Morphological changes were observed after 24-hour culture in neurogenic media, and the transcript levels of the neurogenic markers increased during the prolonged induction period. Moreover, oPB-MSCs expressed the cellular prion protein gene (PRNP), which was up-regulated during neurogenesis.

Conclusions

This study describes for the first time the isolation and characterization of oPB-MSCs. Albeit some variability was observed between animals, these cells retained their capacity to differentiate into mesenchymal lineages and to transdifferentiate into neuron-like cells in vitro. Therefore, oPB-MSCs could serve as a valuable tool for biomedical research in fields including orthopaedics or prion diseases.

Keywords:
Sheep; Mesenchymal stem cell; Peripheral blood; Neurogenesis