Blood type and breed-associated differences in cell marker expression on equine bone marrow-derived mesenchymal stem cells including major histocompatibility complex class II antigen expression.
Abstract: As the search for an immune privileged allogeneic donor mesenchymal stem cell (MSC) line continues in equine medicine, the characterization of the cells between different sources becomes important. Our research seeks to more clearly define the MSC marker expression of different equine MSC donors. The bone marrow-derived MSCs from two equine breeds and different blood donor-types were compared over successive culture passages to determine the differential expression of important antigens. Eighteen Thoroughbreds and 18 Standardbreds, including 8 blood donor (erythrocyte Aa, Ca, and Qa antigen negative) horses, were evaluated. Bone marrow was taken from each horse for isolation and culture of MSCs. Samples from passages 2, 4, 6, and 8 were labelled and evaluated by flow cytometry. The cell surface expression of CD11a/18, CD44, CD90 and MHC class II antigens were assessed. Trilineage assays for differentiation into adipogenic, chondrogenic and osteogenic lines were performed to verify characterization of the cells as MSCs. There were significant differences in mesenchymal stem cell marker expression between breeds and blood antigen-type groups over time. Standardbred horses showed a significantly lower expression of MHC class II than did Thoroughbred horses at passages 2, 4 and 6. CD90 was significantly higher in universal blood donor Standardbreds as compared to non-blood donor Standardbreds over all time points. All MSC samples showed high expression of CD44 and low expression of CD11a/18. Universal blood donor- type Standardbred MSCs from passages 2-4 show the most ideal antigen expression pattern of the horses and passages that we characterized for use as a single treatment of donor bone marrow-derived MSCs. Further work is needed to determine the significance of this differential expression along with the effect of the expression of MHC I on equine bone marrow-derived MSCs.
Publication Date: 2019-11-20 PubMed ID: 31747418PubMed Central: PMC6867698DOI: 10.1371/journal.pone.0225161Google Scholar: Lookup
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Summary
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The research investigates the differences in cell marker expression of mesenchymal stem cells (MSC) derived from the bone marrow of two different horse breeds and different blood type donors, focusing on their expression of major histocompatibility complex (MHC) class II antigens. The study found breed and blood type-associated differences in cell marker expression over time, specifically highlighting universal blood donor type Standardbred horse’s MSCs as ideal for use as donor cells for single treatment bone marrow-derived MSCs.
Introduction
- The study is a part of ongoing research in equine medicine to identify an immune privileged allogeneic donor for mesenchymal stem cells (MSCs).
- Researchers are interested in understanding cell marker expression by different equine MSC donors, as these markers can heavily influence the effectiveness and compatibility of the resulting treatments.
Methodology
- The bone marrow from two different horse breeds (18 Thoroughbreds and 18 Standardbreds), including 8 blood donor horses negative for erythrocyte antigens Aa, Ca, and Qa, was collected for this study.
- The bone marrow was cultured to produce MSCs which were then evaluated at specific culture passages (2, 4, 6, and 8) using flow cytometry.
- Assessment of cell surface expression of CD11a/18, CD44, CD90, and MHC class II antigens was done.
- Trilineage assays were also performed to verify the characterization of cultured cells as MSCs through their capacity to differentiate into adipogenic, chondrogenic, and osteogenic lines.
Results
- There were notable differences in MSC marker expression over time between Standardbred and Thoroughbred horses and among different blood antigen-type groups.
- The researchers found that MSCs from Standardbred horses showed significantly lower MHC class II expression than those from Thoroughbred horses at passages 2, 4, and 6.
- CD90 expression was significantly higher in MSCs from universal blood donor Standardbreds compared to non-blood donor Standardbreds across all time points.
- All MSC samples exhibited high CD44 expression and low CD11a/18 expression.
Conclusions and Further Research
- The findings indicate that the MSCs from universal blood donor type Standardbred horses, particularly from passages 2-4, showed the most ideal antigen expression pattern for use as donor bone marrow-derived MSCs.
- However, more research is needed to understand the biological and clinical implications of these differences in cell marker expressions, especially the effect of MHC I expression on equine bone marrow-derived MSCs.
Cite This Article
APA
Kamm JL, Parlane NA, Riley CB, Gee EK, Dittmer KE, McIlwraith CW.
(2019).
Blood type and breed-associated differences in cell marker expression on equine bone marrow-derived mesenchymal stem cells including major histocompatibility complex class II antigen expression.
PLoS One, 14(11), e0225161.
https://doi.org/10.1371/journal.pone.0225161 Publication
Researcher Affiliations
- Massey University, School of Veterinary Science, Massey University, Palmerston North, New Zealand.
- Veterinary Associates, Karaka, Auckland, New Zealand.
- AgResearch, Hopkirk Research Institute, Massey University, Palmerston North, New Zealand.
- Massey University, School of Veterinary Science, Massey University, Palmerston North, New Zealand.
- Massey University, School of Veterinary Science, Massey University, Palmerston North, New Zealand.
- Massey University, School of Veterinary Science, Massey University, Palmerston North, New Zealand.
- Massey University, School of Veterinary Science, Massey University, Palmerston North, New Zealand.
- Colorado State University, Orthopaedic Research Center, Fort Collins, Colorado, United States of America.
MeSH Terms
- Animals
- Biomarkers / blood
- Bone Marrow Cells / metabolism
- Cells, Cultured
- Histocompatibility Antigens / genetics
- Histocompatibility Antigens / metabolism
- Horses / blood
- Horses / genetics
- Hyaluronan Receptors / genetics
- Hyaluronan Receptors / metabolism
- Mesenchymal Stem Cells / metabolism
- Thy-1 Antigens / genetics
- Thy-1 Antigens / metabolism
- Veterinary Medicine / methods
Conflict of Interest Statement
I have read the journal's policy and the authors of this manuscript have the following competing interests: C. Wayne McIlwraith and Lacy Kamm are directors and owners of Advanced Regenerative Therapies, New Zealand. This does not alter our adherence to PLOS ONE policies on sharing data and materials.
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