Shared senior authorship.
Successful isolation of equine mesenchymal stromal cells from cryopreserved umbilical cord blood-derived mononuclear cell fractions
Article first published online: 4 DEC 2012
© 2012 EVJ Ltd
Equine Veterinary Journal
Volume 45, Issue 4, pages 518–522, July 2013
How to Cite
De Schauwer, C., van de Walle, G. R., Piepers, S., Hoogewijs, M. K., Govaere, J. L. J., Meyer, E. and van Soom, A. (2013), Successful isolation of equine mesenchymal stromal cells from cryopreserved umbilical cord blood-derived mononuclear cell fractions. Equine Veterinary Journal, 45: 518–522. doi: 10.1111/evj.12003
- Issue published online: 6 JUN 2013
- Article first published online: 4 DEC 2012
- Accepted manuscript online: 12 OCT 2012 06:05AM EST
- Manuscript Accepted: 3 OCT 2012
- Manuscript Received: 11 APR 2012
- mesenchymal stromal cells;
- mononuclear cell fraction
Reasons for performing study
The therapeutic potential of mesenchymal stromal cells for cellular therapy has generated increasing interest in human as well as veterinary medicine. Considerable research has been performed on the cryopreservation of expanded mesenchymal stromal cells, but little information is available on the cryopreservation of the original mononuclear cell fraction.
The present study describes a protocol to expand equine mesenchymal stromal cells after cryopreserving the mononuclear cells of umbilical cord blood.
To this end, mononuclear cells were isolated from 7 umbilical cord blood samples and cryopreserved at a concentration of 1–2 × 109 cells/l cold freezing solution. Cells were cryopreserved and kept frozen for at least 6 months before thawing. Frozen cryotubes were thawed in a 37°C water bath. Putative equine mesenchymal stromal cells were immunophenotyped using multicolour flow cytometry based on a selected 9 marker panel.
Average cell viability upon thawing was 98.7 ± 0.6%. In 6 out of 7 samples, adherent spindle-shaped cell colonies were observed within 9.0 ± 2.6 days and attained 80% confluency at 12.3 ± 3.9 days. After 3 passages, putative equine mesenchymal stromal cells were successfully immunophenotyped as CD29, CD44 and CD90 positive, and CD45, CD73, CD79α, CD105, MHC II and monocyte-marker negative.
Conclusions and potential relevance
Equine mesenchymal stromal cells can be cultured after cryopreservation of the isolated mononuclear cells, a time- as well as cost-efficient approach in equine regenerative medicine.