FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / PLoS One / Post-thaw non-cultured and post-thaw cultured equine cord bl...
PloS one2014; 9(12); e113615; doi: 10.1371/journal.pone.0113615

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

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.
Get Full Text
Publication Date: 2014-12-01 PubMed ID: 25438145PubMed Central: PMC4249887DOI: 10.1371/journal.pone.0113615Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • 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 found that the immunosuppressive qualities of frozen equine cord blood mesenchymal stromal cells (MSC) remain intact even after being thawed and immediately used in suppressive tests, with no need for further culturing post-thaw.

Context and Hypothesis

  • The research was built around mesenchymal stromal cells (MSC) obtained from equine cord blood. These cells are known for their potential immunomodulatory characteristics – this means they can modify the immune response or the functioning of the immune system.
  • A common method to store these cells is cryopreservation or freezing them for later use. However, prior studies have shown that when these cells are thawed, they experience a phase of slowed proliferation or ‘lag-phase’.
  • The researchers hypothesized that there would be no difference in the ability of these thawed cells to suppress lymphocyte proliferation, regardless of whether they were used immediately after thawing or allowed to undergo post-thaw culture time before experimental inclusion.

Methodology

  • The team compared cryopreserved cord blood mesenchymal stromal cells (CB-MSC) obtained from five unrelated foals.
  • These cells were subjected to two-way mixed lymphocyte reaction (MLR) assays, which evaluate the interaction of different immune cells and the reaction that ensues.
  • For each of the five cell cultures, separate MLR assays were performed on MSC that were allowed to culture for five days after being thawed and on cells that were included in the MLR assay immediately post-thawing.

Findings

  • The study found that there was no significant difference in the ability of the CB-MSC to suppress lymphocyte proliferation, irrespective of whether they had undergone post-thaw culture or were not cultured post-thaw.
  • There was also no significant difference between the immunosuppressive properties of MSC obtained from five different donors, implying a lack of inter-donor variability.

Implications

  • The findings suggest that cryopreserved CB-MSC could be immediately usable upon thawing in a clinical setting. This would eliminate the need for further cell culture, making the cells more readily available.
  • The implications of this finding could extend to the usage of these cells at third-party locations without the need for specialized cell culture equipment or expertise, thus expediting their use in various research and clinical settings.

Cite This Article

APA
Williams LB, Tessier L, Koenig JB, Koch TG. (2014). Post-thaw non-cultured and post-thaw cultured equine cord blood mesenchymal stromal cells equally suppress lymphocyte proliferation in vitro. PLoS One, 9(12), e113615. https://doi.org/10.1371/journal.pone.0113615

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 9
Issue: 12
Pages: e113615
PII: e113615

Researcher Affiliations

Williams, Lynn B
  • Department of Clinical Studies, University of Guelph, Guelph, Ontario, Canada.
Tessier, Laurence
  • Department of Biomedical Science, University of Guelph, Guelph, Ontario, Canada.
Koenig, Judith B
  • Department of Clinical Studies, University of Guelph, Guelph, Ontario, Canada.
Koch, Thomas G
  • Department of Biomedical Science, University of Guelph, Guelph, Ontario, Canada; Department of Clinical Studies, Orthopaedic Research Lab, Aarhus University, Copenhagen, Denmark.

MeSH Terms

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

Conflict of Interest Statement

TGK acts in a volunteer capacity as non-executive Director, Scientific Affairs (ex officio) of eQcell therapies Inc., Aurora, Ontario, Canada, a company for which TGK's research laboratory provides equine stem cell isolation and storage services. TGK holds a minor non-controlling share in eQcell therapies Inc. LBW, LT and JBK declare no competing interests. This does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials.

References

This article includes 29 references
  1. Sutton EJ, Boddington SE, Nedopil AJ, Henning TD, Demos SG, Baehner R, Sennino B, Lu Y, Daldrup-Link HE. An optical imaging method to monitor stem cell migration in a model of immune-mediated arthritis.. Opt Express 2009 Dec 21;17(26):24403-13.
    pmc: PMC2888495pubmed: 20052149doi: 10.1364/oe.17.024403google scholar: lookup
  2. Toupet K, Maumus M, Peyrafitte JA, Bourin P, van Lent PL, Ferreira R, Orsetti B, Pirot N, Casteilla L, Jorgensen C, Noël D. Long-term detection of human adipose-derived mesenchymal stem cells after intraarticular injection in SCID mice.. Arthritis Rheum 2013 Jul;65(7):1786-94.
    pubmed: 23553439doi: 10.1002/art.37960google scholar: lookup
  3. Pigott JH, Ishihara A, Wellman ML, Russell DS, Bertone AL. Investigation of the immune response to autologous, allogeneic, and xenogeneic mesenchymal stem cells after intra-articular injection in horses.. Vet Immunol Immunopathol 2013 Nov 15;156(1-2):99-106.
    pubmed: 24094688doi: 10.1016/j.vetimm.2013.09.003google scholar: lookup
  4. Di Nicola M, Carlo-Stella C, Magni M, Milanesi M, Longoni PD, Matteucci P, Grisanti S, Gianni AM. Human bone marrow stromal cells suppress T-lymphocyte proliferation induced by cellular or nonspecific mitogenic stimuli.. Blood 2002 May 15;99(10):3838-43.
    pubmed: 11986244doi: 10.1182/blood.v99.10.3838google scholar: lookup
  5. Leri A, Kajstura J, Anversa P. Cardiac stem cells and mechanisms of myocardial regeneration.. Physiol Rev 2005 Oct;85(4):1373-416.
    pubmed: 16183916doi: 10.1152/physrev.00013.2005google scholar: lookup
  6. Terada N, Hamazaki T, Oka M, Hoki M, Mastalerz DM, Nakano Y, Meyer EM, Morel L, Petersen BE, Scott EW. Bone marrow cells adopt the phenotype of other cells by spontaneous cell fusion.. Nature 2002 Apr 4;416(6880):542-5.
    pubmed: 11932747doi: 10.1038/nature730google scholar: lookup
  7. Ying QL, Nichols J, Evans EP, Smith AG. Changing potency by spontaneous fusion.. Nature 2002 Apr 4;416(6880):545-8.
    pubmed: 11932748doi: 10.1038/nature729google scholar: lookup
  8. Gallagher KL, Benfey PN. Not just another hole in the wall: understanding intercellular protein trafficking.. Genes Dev 2005 Jan 15;19(2):189-95.
    pubmed: 15655108doi: 10.1101/gad.1271005google scholar: lookup
  9. Rustom A, Saffrich R, Markovic I, Walther P, Gerdes HH. Nanotubular highways for intercellular organelle transport.. Science 2004 Feb 13;303(5660):1007-10.
    pubmed: 14963329doi: 10.1126/science.1093133google scholar: lookup
  10. Park JE, Seo YK, Yoon HH, Kim CW, Park JK, Jeon S. Electromagnetic fields induce neural differentiation of human bone marrow derived mesenchymal stem cells via ROS mediated EGFR activation.. Neurochem Int 2013 Mar;62(4):418-24.
    pubmed: 23411410doi: 10.1016/j.neuint.2013.02.002google scholar: lookup
  11. Torrente D, Avila MF, Cabezas R, Morales L, Gonzalez J, Samudio I, Barreto GE. Paracrine factors of human mesenchymal stem cells increase wound closure and reduce reactive oxygen species production in a traumatic brain injury in vitro model.. Hum Exp Toxicol 2014 Jul;33(7):673-84.
    pubmed: 24178889doi: 10.1177/0960327113509659google scholar: lookup
  12. Nyamandi VZ, Johnsen VL, Hughey CC, Hittel DS, Khan A, Newell C, Shearer J. Enhanced stem cell engraftment and modulation of hepatic reactive oxygen species production in diet-induced obesity.. Obesity (Silver Spring) 2014 Mar;22(3):721-9.
    pubmed: 23894091doi: 10.1002/oby.20580google scholar: lookup
  13. Quintanilha LF, Takami T, Hirose Y, Fujisawa K, Murata Y, Yamamoto N, Goldenberg RC, Terai S, Sakaida I. Canine mesenchymal stem cells show antioxidant properties against thioacetamide-induced liver injury in vitro and in vivo.. Hepatol Res 2014 Oct;44(10):E206-17.
    pubmed: 23889977doi: 10.1111/hepr.12204google scholar: lookup
  14. Keating A. How do mesenchymal stromal cells suppress T cells?. Cell Stem Cell 2008 Feb 7;2(2):106-8.
    pubmed: 18371428doi: 10.1016/j.stem.2008.01.007google scholar: lookup
  15. Zhou Y, Day A, Haykal S, Keating A, Waddell TK. Mesenchymal stromal cells augment CD4+ and CD8+ T-cell proliferation through a CCL2 pathway.. Cytotherapy 2013 Oct;15(10):1195-207.
    pubmed: 23845188doi: 10.1016/j.jcyt.2013.05.009google scholar: lookup
  16. Carrade DD, Lame MW, Kent MS, Clark KC, Walker NJ, Borjesson DL. Comparative Analysis of the Immunomodulatory Properties of Equine Adult-Derived Mesenchymal Stem Cells().. Cell Med 2012;4(1):1-11.
    pmc: PMC3495591pubmed: 23152950doi: 10.3727/215517912x647217google scholar: lookup
  17. nTessier L. Isolation, immunophenotyping and lymphocyte suppressive properties of equine cord blood-derived mesenchymal stromal cells. 2013.
  18. Kang JG, Park SB, Seo MS, Kim HS, Chae JS, Kang KS. Characterization and clinical application of mesenchymal stem cells from equine umbilical cord blood.. J Vet Sci 2013;14(3):367-71.
    pmc: PMC3788164pubmed: 23820166doi: 10.4142/jvs.2013.14.3.367google scholar: lookup
  19. Lehle K, Hoenicka M, Jacobs VR, Schmid FX, Birnbaum DE. Cryopreservation of human endothelial cells for vascular tissue engineering.. Cryobiology 2005 Apr;50(2):154-61.
    pubmed: 15843005doi: 10.1016/j.cryobiol.2004.12.006google scholar: lookup
  20. Koch TG, Heerkens T, Thomsen PD, Betts DH. Isolation of mesenchymal stem cells from equine umbilical cord blood.. BMC Biotechnol 2007 May 30;7:26.
    pmc: PMC1904213pubmed: 17537254doi: 10.1186/1472-6750-7-26google scholar: lookup
  21. Koch TG, Thomsen PD, Betts DH. Improved isolation protocol for equine cord blood-derived mesenchymal stromal cells.. Cytotherapy 2009;11(4):443-7.
    pubmed: 19513899doi: 10.1080/14653240902887259google scholar: lookup
  22. Frisbie D. Synovial joint biology and pathobiology. 2012.
  23. Osteoarthritis CJ (2003) In: Ross M, Dyson Seditors. In: Diagnosis and management of lameness in the horse. St. Louis: Saunders; pp.572–591.
  24. Shirtliff ME, Mader JT. Acute septic arthritis.. Clin Microbiol Rev 2002 Oct;15(4):527-44.
    pmc: PMC126863pubmed: 12364368doi: 10.1128/cmr.15.4.527-544.2002google scholar: lookup
  25. Roy S, Bhawan J. Ultrastructure of articular cartilage in pyogenic arthritis.. Arch Pathol 1975 Jan;99(1):44-7.
    pubmed: 1111494
  26. Carrade DD, Owens SD, Galuppo LD, Vidal MA, Ferraro GL, Librach F, Buerchler S, Friedman MS, Walker NJ, Borjesson DL. Clinicopathologic findings following intra-articular injection of autologous and allogeneic placentally derived equine mesenchymal stem cells in horses.. Cytotherapy 2011 Apr;13(4):419-30.
    pubmed: 21105841doi: 10.3109/14653249.2010.536213google scholar: lookup
  27. Ricco S, Renzi S, Del Bue M, Conti V, Merli E, Ramoni R, Lucarelli E, Gnudi G, Ferrari M, Grolli S. Allogeneic adipose tissue-derived mesenchymal stem cells in combination with platelet rich plasma are safe and effective in the therapy of superficial digital flexor tendonitis in the horse.. Int J Immunopathol Pharmacol 2013 Jan-Mar;26(1 Suppl):61-8.
    pubmed: 24046950doi: 10.1177/03946320130260s108google scholar: lookup
  28. François M, Copland IB, Yuan S, Romieu-Mourez R, Waller EK, Galipeau J. Cryopreserved mesenchymal stromal cells display impaired immunosuppressive properties as a result of heat-shock response and impaired interferon-γ licensing.. Cytotherapy 2012 Feb;14(2):147-52.
    pmc: PMC3279133pubmed: 22029655doi: 10.3109/14653249.2011.623691google scholar: lookup
  29. Xu X, Liu Y, Cui Z, Wei Y, Zhang L. Effects of osmotic and cold shock on adherent human mesenchymal stem cells during cryopreservation.. J Biotechnol 2012 Dec 31;162(2-3):224-31.
    pubmed: 22989486doi: 10.1016/j.jbiotec.2012.09.004google scholar: lookup

Citations

This article has been cited 6 times.
  1. Jammes M, Contentin R, Cassé F, Galéra P. Equine osteoarthritis: Strategies to enhance mesenchymal stromal cell-based acellular therapies.. Front Vet Sci 2023;10:1115774.
    doi: 10.3389/fvets.2023.1115774pubmed: 36846261google scholar: lookup
  2. Mund SJK, MacPhee DJ, Campbell J, Honaramooz A, Wobeser B, Barber SM. Macroscopic, Histologic, and Immunomodulatory Response of Limb Wounds Following Intravenous Allogeneic Cord Blood-Derived Multipotent Mesenchymal Stromal Cell Therapy in Horses.. Cells 2021 Nov 1;10(11).
    doi: 10.3390/cells10112972pubmed: 34831196google scholar: lookup
  3. Nguyen MQ, Bui HTH, Tuyet ANT, Nhung TTH, Hoang DM, Liem NT, Hoang VT. Comparative Bioactivity Analysis for Off-the-Shelf and Culture-Rescued Umbilical Cord-Derived Mesenchymal Stem/Stromal Cells in a Xeno- and Serum-Free Culture System.. Cell Transplant 2021 Jan-Dec;30:9636897211039441.
    doi: 10.1177/09636897211039441pubmed: 34538123google scholar: lookup
  4. Mund SJK, Kawamura E, Awang-Junaidi AH, Campbell J, Wobeser B, MacPhee DJ, Honaramooz A, Barber S. Homing and Engraftment of Intravenously Administered Equine Cord Blood-Derived Multipotent Mesenchymal Stromal Cells to Surgically Created Cutaneous Wound in Horses: A Pilot Project.. Cells 2020 May 8;9(5).
    doi: 10.3390/cells9051162pubmed: 32397125google scholar: lookup
  5. Williams LB, Co C, Koenig JB, Tse C, Lindsay E, Koch TG. Response to Intravenous Allogeneic Equine Cord Blood-Derived Mesenchymal Stromal Cells Administered from Chilled or Frozen State in Serum and Protein-Free Media.. Front Vet Sci 2016;3:56.
    doi: 10.3389/fvets.2016.00056pubmed: 27500136google scholar: lookup
  6. Williams LB, Russell KA, Koenig JB, Koch TG. Aspiration, but not injection, decreases cultured equine mesenchymal stromal cell viability.. BMC Vet Res 2016 Mar 7;12:45.
    doi: 10.1186/s12917-016-0671-2pubmed: 26952099google scholar: lookup
Subscribe to our newsletter
Join 99,359 horse owners like you who receive our email updates on horse health and nutrition.