Comparative immunophenotyping of equine multipotent mesenchymal stromal cells: an approach toward a standardized definition.
Abstract: Horses are an approved large animal model for therapies of the musculoskeletal system. Especially for tendon disease where cell-based therapy is commonly used in equine patients, the translation of achieved results to human medicine would be a great accomplishment. Immunophenotyping of equine mesenchymal stromal cells (MSCs) remains the last obstacle to meet the criteria of the International Society for Cellular Therapy (ISCT) definition of human MSCs. Therefore, the surface antigen expression of CD 29, CD 44, CD 73, CD 90, CD 105, CD 14, CD 34, CD 45, CD 79α, and MHC II in equine MSCs from adipose tissue, bone marrow, umbilical cord blood, umbilical cord tissue, and tendon tissue was analyzed using flow cytometry. Isolated cells from the different sources and donors varied in their expression pattern of MSC-defining antigens. In particular, CD 90 and 105 showed most heterogeneity. However, cells from all samples were robustly positive for CD 29 and CD 44, while being mostly negative for CD 73 and the exclusion markers CD 14, CD 34, CD 45, CD 79α and MHC II. Furthermore, it was evident that enzymes used for cell detachment after in vitro-culture affected the detection of antigen expression. These results emphasize the need of standardization of MSC isolation, culturing, and harvesting techniques. As the equine MSCs did not meet all criteria the ISCT defined for human MSCs, further investigations for a better characterization of the cell type should be conducted.
© 2014 International Society for Advancement of Cytometry.
Publication Date: 2014-06-03 PubMed ID: 24894974DOI: 10.1002/cyto.a.22491Google Scholar: Lookup
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The researchers in this article analyze the surface antigen expression of specific markers in different equine mesenchymal stromal cells (MSCs) to standardize their definition in line with those of human MSCs by the International Society for Cellular Therapy (ISCT). They found that the cells varied significantly in their antigen expression depending on their sources and donors, and that the use of enzymes in vitro affected antigen detection.
Objective of the Research
- The main goal of this study is to understand and categorize the immune characteristics or immunophenotyping of equine mesenchymal stromal cells (MSCs).
- The researchers also aimed to standardize how these cells are defined, in alignment with the criteria established by the International Society for Cellular Therapy (ISCT) for human MSCs.
Significance of the Study
- Horses are used as a large animal model for musculoskeletal system therapies. Particularly in the case of tendon diseases, cell-based therapies have been successfully employed in equine patients.
- Translating these results to human medicine could prove highly beneficial. To achieve this, however, a clear understanding and definition of equine MSCs is required.
Methodology and Findings
- The expression of specific surface antigens — specifically CD 29, CD 44, CD 73, CD 90, CD 105, CD 14, CD 34, CD 45, CD 79α, and MHC II — in equine MSCs derived from adipose tissue, bone marrow, umbilical cord blood, umbilical cord tissue, and tendon tissue was examined using flow cytometry.
- The researchers found that the isolated cells from the different sources and donors varied in their expression pattern of these MSC-defining antigens. In particular, CD 90 and 105 displayed the greatest heterogeneity.
- Cells from all the tested samples were mostly positive for CD 29 and CD 44, while being predominantly negative for CD 73 and the exclusion markers CD 14, CD 34, CD 45, CD 79α and MHC II.
- It was also noted that the enzymes used for cell detachment in lab conditions affected the detection of antigen expression.
Conclusion and Recommendations
- The results highlight the need for standardization of procedures involved in the isolation, culturing, and harvesting of MSCs.
- The equine MSCs tested did not meet all the criteria set by the ISCT for human MSCs, suggesting a need for further research to better characterize this cell type.
Cite This Article
APA
Paebst F, Piehler D, Brehm W, Heller S, Schroeck C, Tárnok A, Burk J.
(2014).
Comparative immunophenotyping of equine multipotent mesenchymal stromal cells: an approach toward a standardized definition.
Cytometry A, 85(8), 678-687.
https://doi.org/10.1002/cyto.a.22491 Publication
Researcher Affiliations
- Translational Centre for Regenerative Medicine (TRM), University of Leipzig, Leipzig, Germany; Faculty of Veterinary Medicine, Large Animal Clinic for Surgery, University of Leipzig, Leipzig, Germany.
MeSH Terms
- Animals
- Antibodies / immunology
- Antigens, Surface / metabolism
- Biomarkers / metabolism
- Cell Adhesion
- Cross Reactions
- Female
- Flow Cytometry
- Horses
- Humans
- Immunophenotyping / methods
- Immunophenotyping / standards
- Mesenchymal Stem Cells / cytology
- Mesenchymal Stem Cells / immunology
- Multipotent Stem Cells / cytology
- Multipotent Stem Cells / immunology
- Reference Standards
Citations
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