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Stem Cell Res Ther. 2015 Dec 18;6:253. doi: 10.1186/s13287-015-0250-7.

Tissues from equine cadaver ligaments up to 72 hours of post-mortem: a promising reservoir of stem cells.

Author information

1
Anatomy Unit, FARAH Research Center & Faculty of Veterinary Medicine, University of Liège, Liège, Belgium. mkal.sook@doct.ulg.ac.be.
2
Anatomy Unit, FARAH Research Center & Faculty of Veterinary Medicine, University of Liège, Liège, Belgium. annick.gabriel@ulg.ac.be.
3
Histology Unit, FARAH Research Center & Faculty of Veterinary Medicine, University of Liège, Liège, Belgium. jpiret@ulg.ac.be.
4
Histology Unit, FARAH Research Center & Faculty of Veterinary Medicine, University of Liège, Liège, Belgium. o.waroux@ulg.ac.be.
5
Embryology Unit, FARAH Research Center & Faculty of Veterinary Medicine, University of Liège, Liège, Belgium. ctonus@ulg.ac.be.
6
Embryology Unit, GIGA-Development, Stem Cells and Regenerative Medicine and Faculty of Veterinary Medicine, University of Liège, Liège, Belgium. delphine.connan@ulg.ac.be.
7
FARAH Research Center & Faculty of Veterinary Medicine, University of Liège, Liège, Belgium. etienne.baise@ulg.ac.be.
8
Histology Unit, FARAH Research Center & Faculty of Veterinary Medicine, University of Liège, Liège, Belgium. nadine.antoine@ulg.ac.be.

Abstract

BACKGROUND:

Mesenchymal stem cells (MSCs) harvested from cadaveric tissues represent a promising approach for regenerative medicine. To date, no study has investigated whether viable MSCs could survive in cadaveric tissues from tendon or ligament up to 72 hours of post-mortem. The purpose of the present work was to find out if viable MSCs could survive in cadaveric tissues from adult equine ligaments up to 72 hours of post-mortem, and to assess their ability (i) to remain in an undifferentiated state and (ii) to divide and proliferate in the absence of any specific stimulus.

METHODS:

MSCs were isolated from equine cadaver (EC) suspensory ligaments within 48-72 hours of post-mortem. They were evaluated for viability, proliferation, capacity for tri-lineage differentiation, expression of cell surface markers (CD90, CD105, CD73, CD45), pluripotent transcription factor (OCT-4), stage-specific embryonic antigen-1 (SSEA-1), neuron-specific class III beta-tubulin (TUJ-1), and glial fibrillary acidic protein (GFAP). As well, they were characterized by transmission electron microscope (TEM).

RESULTS:

EC-MSCs were successfully isolated and maintained for 20 passages with high cell viability and proliferation. Phase contrast microscopy revealed that cells with fibroblast-like appearance were predominant in the culture. Differentiation assays proved that EC-MSCs are able to differentiate towards mesodermal lineages (osteogenic, adipogenic, chondrogenic). Flow cytometry analysis demonstrated that EC-MSCs expressed CD90, CD105, and CD73, while being negative for the leukocyte common antigen CD45. Immunofluorescence analysis showed a high percentage of positive cells for OCT-4 and SSEA-1. Surprisingly, in absence of any stimuli, some adherent cells closely resembling neuronal and glial morphology were also observed. Interestingly, our results revealed that approximately 15 % of the cell populations were TUJ-1 positive, whereas GFAP expression was detected in only a few cells. Furthermore, TEM analysis confirmed the stemness of EC-MSCs and identified some cells with a typical neuronal morphology.

CONCLUSIONS:

Our findings raise the prospect that the tissues harvested from equine ligaments up to 72 hours of post-mortem represent an available reservoir of specific stem cells. EC-MSCs could be a promising alternative source for tissue engineering and stem cell therapy in equine medicine.

PMID:
26684484
PMCID:
PMC4683699
DOI:
10.1186/s13287-015-0250-7
[Indexed for MEDLINE]
Free PMC Article

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