Post-thaw non-cultured and post-thaw cultured equine cord blood mesenchymal stromal cells equally suppress lymphocyte proliferation in vitro

PLoS One. 2014 Dec 1;9(12):e113615. doi: 10.1371/journal.pone.0113615. eCollection 2014.

Abstract

Multipotent mesenchymal stromal cells (MSC) are receiving increased attention for their non-progenitor immunomodulatory potential. Cryopreservation is commonly used for long-term storage of MSC. Post-thaw MSC proliferation is associated with a lag-phase in vitro. How this lag-phase affect MSC immunomodulatory properties is unknown. We hypothesized that in vitro there is no difference in lymphocyte suppression potential between quick-thawed cryopreserved equine cord blood (CB) MSC immediately included in mixed lymphocyte reaction (MLR) and same MSC allowed post-thaw culture time prior to inclusion in MLR. Cryopreserved CB-MSC from five unrelated foals were compared using two-way MLR. For each of the five unrelated MSC cultures, paired MLR assays of MSC allowed five days of post-thaw culture and MSC included in MLR assay immediately post-thawing were evaluated. We report no difference in the suppression of lymphocyte proliferation by CB-MSC that had undergone post-thaw culture and MSC not cultured post-thaw (p<0.0001). Also, there was no inter-donor variability between the lymphocyte suppressive properties of MSC harvested from the five different donors (p = 0.13). These findings suggest that cryopreserved CB-MSC may have clinical utility immediately upon thawing. One implication hereof is the possibility of using cryopreserved CB-MSC at third party locations without the need for cell culture equipment or competencies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Proliferation
  • Cells, Cultured
  • Cryopreservation
  • Horses
  • In Vitro Techniques
  • Lymphocyte Culture Test, Mixed / methods*
  • Lymphocytes / cytology*
  • Mesenchymal Stem Cells / immunology*

Grants and funding

These studies were supported by The Danish Council for Independent Research - Council for Technology and Production Sciences (TGK grant #902539; http://ufm.dk/en/research-and-innovation/funding-programmes-for-research-and-innovation/find-danish-funding-programmes/dff-the-danish-council-for-independent-research) and the Equine Guelph Research Foundation (TGK grant #EG 2012 12; http://www.equineguelph.ca/research/index.php). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.