Open Access Highly Accessed Research article

Effect of hypoxia on equine mesenchymal stem cells derived from bone marrow and adipose tissue

Beatriz Ranera1, Ana Rosa Remacha1, Samuel Álvarez-Arguedas1, Antonio Romero2, Francisco José Vázquez2, Pilar Zaragoza1, Inmaculada Martín-Burriel1 and Clementina Rodellar13*

Author Affiliations

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

2 Hospital Veterinario, Facultad de Veterinaria, Universidad de Zaragoza, 50013, Zaragoza, Spain

3 Instituto Aragonés de Ciencias de la Salud (IACS), Zaragoza, 50009, Spain

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BMC Veterinary Research 2012, 8:142  doi:10.1186/1746-6148-8-142

Published: 22 August 2012

Abstract

Background

Mesenchymal stem cells (MSCs) derived from bone marrow (BM-MSCs) and adipose tissue (AT-MSCs) are being applied to equine cell therapy. The physiological environment in which MSCs reside is hypoxic and does not resemble the oxygen level typically used in in vitro culture (20% O2). This work compares the growth kinetics, viability, cell cycle, phenotype and expression of pluripotency markers in both equine BM-MSCs and AT-MSCs at 5% and 20% O2.

Results

At the conclusion of culture, fewer BM-MSCs were obtained in hypoxia than in normoxia as a result of significantly reduced cell division. Hypoxic AT-MSCs proliferated less than normoxic AT-MSCs because of a significantly higher presence of non-viable cells during culture. Flow cytometry analysis revealed that the immunophenotype of both MSCs was maintained in both oxygen conditions. Gene expression analysis using RT-qPCR showed that statistically significant differences were only found for CD49d in BM-MSCs and CD44 in AT-MSCs. Similar gene expression patterns were observed at both 5% and 20% O2 for the remaining surface markers. Equine MSCs expressed the embryonic markers NANOG, OCT4 and SOX2 in both oxygen conditions. Additionally, hypoxic cells tended to display higher expression, which might indicate that hypoxia retains equine MSCs in an undifferentiated state.

Conclusions

Hypoxia attenuates the proliferative capacity of equine MSCs, but does not affect the phenotype and seems to keep them more undifferentiated than normoxic MSCs.

Keywords:
Hypoxia; Horse; AT-MSC; BM-MSC; Characterisation