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Stem cells and development2021; 31(1-2); 18-25; doi: 10.1089/scd.2021.0189

Steps Toward Standardized In Vitro Assessment of Immunomodulatory Equine Mesenchymal Stromal Cells Before Clinical Application.

Abstract: Inflammation-associated disorders are significant causes of morbidity in horses. Equine single-donor mesenchymal stromal cells (sdMSCs) hold promise as cell-therapy candidates due to their secretory nonprogenitor functions. This has been demonstrated by mononuclear cell suppression assays (MSAs) showing that sdMSCs are blood mononuclear cell (BMC) suppressive in vitro. sdMSCs derived from umbilical cord blood are of clinical interest due to their ease of procurement, multipotency, and immunomodulatory ability. Due to the inherent donor-to-donor heterogeneity of MSCs, the development of robust and easily deployable methods of potency assessment is critical for improving MSCs' predictability in treating inflammatory diseases. This study focuses on the development of robust in vitro potency assays and the assessment of potential sdMSC therapeutic end products generated from pooled sdMSCs (pMSCs). We hypothesized that, compared to MSA using only one donor, MSA using pooled BMCs (pBMCs) is a more robust sdMSC potency assay due to reduced donor BMC heterogeneity. pBMCs were generated by pooling equine BMCs isolated from peripheral blood of five donors in equal ratios. pBMCs were labeled with carboxyfluorescein succinimidyl ester (CFSE) and stored in liquid nitrogen until use. Similarly, pooling sdMSCs from multiple equine donors in equal ratios generated pMSCs. sdMSC cultures were assessed with pBMCs in MSA using Bromodeoxyuridine ELISA and CFSE. Proliferation assessment of BMCs from individual donors revealed varied responses to concanavalin A (ConA) stimulation. MSA using BMCs from single donors further demonstrated BMC donor variability. Utilizing this assay, we have also found that the immunosuppressive potencies of pMSCs are at least equal, if not more, than the calculated mean of individual cultures. MSA based on pBMCs provides a consistent and reproducible equine sdMSC potency assay. This knowledge could be used in production monitoring of cellular potency and as release criteria before clinical use.
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Publication Date: 2021-12-03 PubMed ID: 34779250PubMed Central: PMC8792491DOI: 10.1089/scd.2021.0189Google 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.

Equine single-donor mesenchymal stromal cells (sdMSCs) show potential for cell-therapy application because of their immunomodulatory properties. However, variations among donors complicate the standardization of potency assessments, affecting the predictability of these cells in treating inflammatory diseases. The research paper suggests an alternative method, using pooled blood mononuclear cells (pBMCs) instead of individual donor cells, for more consistent in vitro potency assays.

Objective and Hypothesis

  • The study’s objective was to develop robust and easily deployable in vitro methods for assessing sdMSCs’ potency before using them in clinical treatments for inflammation-associated diseases in horses.
  • The primary hypothesis was that using pooled blood mononuclear cells (pBMCs) — a mixture collected from multiple donors — in mononuclear cell suppression assays (MSAs) would yield more robust results than using cells from a single donor, due to the reduced variability inherent in the former method.

Methods

  • The researchers created pBMCs by combining blood mononuclear cells isolated from the peripheral blood of five different donors in equal ratios.
  • Similarly, to produce pooled sdMSCs (pMSCs), they mixed sdMSCs from multiple donors, also in equal ratios.
  • Both pBMCs and pMSCs were labeled with carboxyfluorescein succinimidyl ester (CFSE) and stored in liquid nitrogen.
  • Various assays assessed the suppressive potential of these cells, including mononuclear cell suppression assays (MSAs), Bromodeoxyuridine ELISAs, and tests using Concanavalin A stimulation.

Findings

  • The researchers found varied responses to Concanavalin A stimulation in the proliferation assessment of BMCs from individual donors, suggesting donor variability could complicate potency assessments.
  • The use of pBMCs in the MSAs allowed for consistency and reproducibility in evaluating the potency of equine sdMSCs.
  • In addition, testing showed the immunosuppressive potencies of pMSCs were at least on par, if not better than, the calculated mean of individual cultures.

Significance

  • This study not only suggests a solution to the variability challenge in sdMSC potency assessments but also provides a reliable procedure to test future cellular products before clinical application. This will enhance treatment predictability for inflammation-associated disorders in horses.
  • The researchers suggest this potency-assessment method could be used for production control and as part of the release criteria before clinical use.

Cite This Article

APA
Lee OJ, Koch TG. (2021). Steps Toward Standardized In Vitro Assessment of Immunomodulatory Equine Mesenchymal Stromal Cells Before Clinical Application. Stem Cells Dev, 31(1-2), 18-25. https://doi.org/10.1089/scd.2021.0189

Publication

Stem cells and development
ISSN: 1557-8534
NlmUniqueID: 101197107
Country: United States
Language: English
Volume: 31
Issue: 1-2
Pages: 18-25

Researcher Affiliations

Lee, Olivia J
  • Department of Biomedical Sciences, University of Guelph, Guelph, Canada.
Koch, Thomas G
  • Department of Biomedical Sciences, University of Guelph, Guelph, Canada.

MeSH Terms

  • Animals
  • Cell Proliferation / physiology
  • Cells, Cultured
  • Fetal Blood
  • Horses
  • Immunomodulation
  • Leukocytes, Mononuclear
  • Mesenchymal Stem Cells

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