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PloS one2015; 10(4); e0122954; doi: 10.1371/journal.pone.0122954

Phenotypic and immunomodulatory properties of equine cord blood-derived mesenchymal stromal cells.

Abstract: Multipotent mesenchymal stromal cells (MSC) have attracted interest for their cytotherapeutic potential, partly due to their immunomodulatory abilities. The aim of this study was to test the robustness of our equine cord blood (CB) MSC isolation protocol, to characterize the CB-MSC before and after cryopreservation, and to evaluate their immunosuppressive phenotype. We hypothesized that MSC can be consistently isolated from equine CB, have unique and reproducible marker expression and in vitro suppress lymphoproliferation. Preliminary investigation of constitutive cytoplasmic Toll-like receptor (TLR) 3 and 4 expression was also preformed due to their possible association with anti- or pro-inflammatory MSC phenotypes, respectively. Surface markers were assessed for antigen and mRNA expression by flow cytometry and quantitative polymerase chain reaction (qPCR). Immunomodulatory properties were evaluated in mixed lymphocyte reaction assays, and TLR3 and TLR4 expression were measured by qPCR and immunocytochemistry (ICC). CB-MSC were isolated from each off nine cord blood samples. CB-MSC highly expressed CD29, CD44, CD90, and lacked or had low expression of major histocompatibility complex (MHC) class I, MHC-II, CD4, CD8, CD11a/18 and CD73 before and after cryopreservation. CB-MSC suppressed in vitro lymphoproliferation and constitutively expressed TLR4. Our findings confirmed CB as a reliable MSC source, provides an association of surface marker phenotype and mRNA expression and suggest anti-inflammatory properties of CB-MSC. The relationship between TLRs and lymphocyte function warrants further investigation.
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Publication Date: 2015-04-22 PubMed ID: 25902064PubMed Central: PMC4406608DOI: 10.1371/journal.pone.0122954Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

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 explores the potential of mesenchymal stromal cells (MSC) derived from equine cord blood for immune suppression and healing applications. The study also delves into the stability of these cells after cryopreservation and their characteristic features.

Objective and Hypothesis

  • The main goal of this research was to investigate the effectiveness of the existing protocol for isolating MSC from equine cord blood and to further study the behavior and features of these cells both before and after cryopreservation.
  • The researchers hypothesized that MSC could be reliably obtained from equine cord blood, and that these cells would have a unique and reproducible marker expression. They also expected that, under in vitro conditions, these cells would inhibit the proliferation of lymphocytes.

Experiments and Methods Used

  • The study included an initial exploration of Toll-like receptors (TLR) 3 and 4, as there could be a potential relationship between these receptors and the anti-inflammatory or pro-inflammatory phenotypes of MSC.
  • Surface markers on the MSC were studied both in terms of antigen and mRNA expression using flow cytometry and quantitative polymerase chain reaction (qPCR) methods.
  • The researchers also performed mixed lymphocyte reaction assays to better understand the immunomodulatory properties of these cells.
  • The levels of expression of TLR3 and TLR4 were assessed using qPCR and immunocytochemistry (ICC).

Results

  • The researchers successfully isolated CB-MSC from each of the nine samples of cord blood used in the study.
  • The CB-MSC expressed high levels of CD29, CD44, and CD90 markers, and either lacked or expressed very low levels of other markers such as MHC class I, MHC-II, CD4, CD8, CD11a/18 and CD73 both before and after cryopreservation.
  • CB-MSC demonstrated the ability to suppress the proliferation of lymphocytes under in vitro conditions and consistently expressed TLR4.

Conclusion

  • The study findings affirmed the viability of cord blood as a dependable source for MSC and displayed a correlation of surface marker phenotype with mRNA expression.
  • The study suggests that the CB-MSC possess anti-inflammatory properties.
  • Further investigations are recommended to elaborate on the connections between TLRs and lymphocyte function.

Cite This Article

APA
Tessier L, Bienzle D, Williams LB, Koch TG. (2015). Phenotypic and immunomodulatory properties of equine cord blood-derived mesenchymal stromal cells. PLoS One, 10(4), e0122954. https://doi.org/10.1371/journal.pone.0122954

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 10
Issue: 4
Pages: e0122954
PII: e0122954

Researcher Affiliations

Tessier, Laurence
  • Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, Canada.
Bienzle, Dorothee
  • Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, Canada.
Williams, Lynn B
  • Department of Clinical Studies, Ontario Veterinary College, University of Guelph, Guelph, Canada.
Koch, Thomas G
  • Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, Canada; The Orthopaedic Research Lab, Aarhus University, Aarhus, Denmark.

MeSH Terms

  • Animals
  • Antigens, CD / metabolism
  • Biomarkers / blood
  • Cell Proliferation
  • Cell Separation
  • Cryopreservation
  • Fetal Blood / cytology
  • Gene Expression Regulation
  • Horses
  • Humans
  • Immunomodulation
  • Leukocytes / cytology
  • Lymphocytes / cytology
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / immunology
  • Phenotype
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Toll-Like Receptor 3 / genetics
  • Toll-Like Receptor 4 / genetics

Conflict of Interest Statement

T. G. Koch acts in a volunteer capacity as non-executive Director, Scientific Affairs (ex officio) of eQcell therapies Inc., Aurora, Ontario, Canada, a company for which T. G. Koch’s research laboratory provides equine stem cell isolation and storage services. T. G. Koch holds a minor non-controlling share in eQcell therapies Inc. L. Tessier, D. Bienzle and L. B. Williams declare no competing interests. The authors hereby confirm that this does not alter their adherence to all PLOS ONE policies on sharing data and materials, as detailed online in your guide for authors http://www.PLOSone.org/static/editorial.action#competing. This does not alter the authors' adherence to PLOS ONE policies on sharing data and materials.

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Citations

This article has been cited 22 times.
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