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Stem cell research & therapy2015; 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.

Abstract: 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.
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Publication Date: 2015-12-18 PubMed ID: 26684484PubMed Central: PMC4683699DOI: 10.1186/s13287-015-0250-7Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article investigates the viability of obtaining Mesenchymal stem cells (MSCs) from equine cadaver ligaments up to 72 hours after death, with the aim to use these cells in regenerative medicine. The study affirms the possibility and comprehensive potential of these stem cells, offering significant implications for tissue engineering and stem cell therapy in equine medicine.

Objective and Methodology of the Research

  • The article is primarily centered around investigating whether MSCs, which are potentially beneficial for regenerative medicine, can survive in cadaveric tissues from tendon or ligament up to 72 hours post-mortem.
  • The study aims to discover if the stem cells can remain in an undifferentiated state and proliferate without any specific stimulus during this period.
  • To carry out the experiment, the researchers collected MSCs from equine cadaveric suspensory ligaments between 48 to 72 hours after death.
  • The isolated stem cells were then evaluated for viability, proliferation capacity, ability to differentiate into three lines, expression of specific cell surface markers, and pluripotent transcription factor.
  • Moreover, the cells were characterized using a transmission electron microscope for further confirmation.

Key Findings

  • The research was successful in isolating the stem cells from the cadaveric equine tissue and maintaining them for 20 passages with high cell viability and proliferation potential.
  • Cells with fibroblast-like appearance were predominantly found in the culture, and the differentiation assays confirmed that these cells were capable of differentiating into mesodermal lineages.
  • Flow cytometry analysis demonstrated the presence of CD90, CD105, and CD73 markers, which are characteristic identifiers of MSCs in the isolated cells, while they showed negative for leukocyte common antigen CD45.
  • Interestingly, without any specific stimulus, adherent cells resembling neuronal and glial morphologies were identified. It was also found that approximately 15% of cell populations were positive for neuronal marker TUJ-1, while only a few cells expressed GFAP, a glial cell marker.
  • The transmission electron microscope study confirmed the ‘stemness’ of these isolated cells, further validating the potential of these cells.

Conclusions and Implications

  • The findings of the research indicate that equine ligament tissues harvested up to 72 hours post-mortem can serve as a reservoir of specific stem cells.
  • These equine cadaver-derived mesenchymal stem cells (EC-MSCs) could provide a promising alternative source for tissue engineering and stem cell therapy in equine medicine.
  • This research creates a valuable groundwork for using cadaveric tissues in regenerative therapy applications and can open a new perspective in equine veterinary medicine.

Cite This Article

APA
Shikh Alsook MK, Gabriel A, Piret J, Waroux O, Tonus C, Connan D, Baise E, Antoine N. (2015). Tissues from equine cadaver ligaments up to 72 hours of post-mortem: a promising reservoir of stem cells. Stem Cell Res Ther, 6, 253. https://doi.org/10.1186/s13287-015-0250-7

Publication

Stem cell research & therapy
ISSN: 1757-6512
NlmUniqueID: 101527581
Country: England
Language: English
Volume: 6
Pages: 253

Researcher Affiliations

Shikh Alsook, Mohamad Khir
  • Anatomy Unit, FARAH Research Center & Faculty of Veterinary Medicine, University of Liège, Liège, Belgium. mkal.sook@doct.ulg.ac.be.
Gabriel, Annick
  • Anatomy Unit, FARAH Research Center & Faculty of Veterinary Medicine, University of Liège, Liège, Belgium. annick.gabriel@ulg.ac.be.
Piret, Joëlle
  • Histology Unit, FARAH Research Center & Faculty of Veterinary Medicine, University of Liège, Liège, Belgium. jpiret@ulg.ac.be.
Waroux, Olivier
  • Histology Unit, FARAH Research Center & Faculty of Veterinary Medicine, University of Liège, Liège, Belgium. o.waroux@ulg.ac.be.
Tonus, Céline
  • Embryology Unit, FARAH Research Center & Faculty of Veterinary Medicine, University of Liège, Liège, Belgium. ctonus@ulg.ac.be.
Connan, Delphine
  • Embryology Unit, GIGA-Development, Stem Cells and Regenerative Medicine and Faculty of Veterinary Medicine, University of Liège, Liège, Belgium. delphine.connan@ulg.ac.be.
Baise, Etienne
  • FARAH Research Center & Faculty of Veterinary Medicine, University of Liège, Liège, Belgium. etienne.baise@ulg.ac.be.
Antoine, Nadine
  • Histology Unit, FARAH Research Center & Faculty of Veterinary Medicine, University of Liège, Liège, Belgium. nadine.antoine@ulg.ac.be.

MeSH Terms

  • Animals
  • Biomarkers / metabolism
  • Cadaver
  • Cell Culture Techniques / methods
  • Cell Culture Techniques / veterinary
  • Cell Differentiation
  • Cell Proliferation
  • Cell Separation / methods
  • Cell Separation / veterinary
  • Glial Fibrillary Acidic Protein / metabolism
  • Horses / anatomy & histology
  • Horses / metabolism
  • In Vitro Techniques
  • Ligaments / cytology
  • Ligaments / metabolism
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / metabolism
  • Microscopy, Electron, Transmission
  • Octamer Transcription Factor-3 / metabolism
  • Postmortem Changes
  • Time Factors
  • Tissue and Organ Harvesting / methods
  • Tissue and Organ Harvesting / veterinary
  • Tubulin / metabolism

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This article has been cited 11 times.
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