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Home / Equine Research Bank / PeerJ / Evaluation of transport conditions for autologous bone marro...
PeerJ2016; 4; e1773; doi: 10.7717/peerj.1773

Evaluation of transport conditions for autologous bone marrow-derived mesenchymal stromal cells for therapeutic application in horses.

Abstract: Background. Mesenchymal stromal cells (MSCs) are increasingly used for clinical applications in equine patients. For MSC isolation and expansion, a laboratory step is mandatory, after which the cells are sent back to the attending veterinarian. Preserving the biological properties of MSCs during this transport is paramount. The goal of the study was to compare transport-related parameters (transport container, media, temperature, time, cell concentration) that potentially influence characteristics of culture expanded equine MSCs. Methods. The study was arranged in three parts comparing (I) five different transport containers (cryotube, two types of plastic syringes, glass syringe, CellSeal), (II) seven different transport media, four temperatures (4 °C vs. room temperature; -20 °C vs. -80 °C), four time frames (24 h vs. 48 h; 48 h vs. 72 h), and (III) three MSC concentrations (5 × 10(6), 10 × 10(6), 20 × 10(6) MSC/ml). Cell viability (Trypan Blue exclusion; percent and total number viable cell), proliferation and trilineage differentiation capacity were assessed for each test condition. Further, the recovered volume of the suspension was determined in part I. Each condition was evaluated using samples of six horses (n = 6) and differentiation protocols were performed in duplicates. Results. In part I of the study, no significant differences in any of the parameters were found when comparing transport containers at room temperature. The glass syringe was selected for all subsequent evaluations (highest recoverable volume of cell suspension and cell viability). In part II, media, temperatures, or time frames had also no significant influence on cell viability, likely due to the large number of comparisons and small sample size. Highest cell viability was observed using autologous bone marrow supernatant as transport medium, and "transport" at 4 °C for 24 h (70.6% vs. control group 75.3%); this was not significant. Contrary, viability was unacceptably low (<40%) for all freezing protocols at -20 °C or -80 °C, particularly with bone marrow supernatant or plasma and DMSO. In part III, various cell concentrations also had no significant influence on any of the evaluated parameters. Chondrogenic differentiation showed a trend towards being decreased for all transport conditions, compared to control cells. Discussion. In this study, transport conditions were not found to impact viability, proliferation or ability for trilineage differentiation of MSCs, most likely due to the small sample size and large number of comparisons. The unusual low viability after all freezing protocols is in contrast to previous equine studies. Potential causes are differences in the freezing, but also in thawing method. Also, the selected container (glass syringe) may have impacted viability. Future research may be warranted into the possibly negative effect of transport on chondrogenic differentiation.
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Publication Date: 2016-03-22 PubMed ID: 27019778PubMed Central: PMC4806605DOI: 10.7717/peerj.1773Google Scholar: Lookup
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Summary

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This research studied the impact of different transport conditions on the viability of horse bone marrow-derived mesenchymal stromal cells (MSCs) and found that, despite common assumptions, the cells demonstrated remarkable resilience to a variety of conditions.

Research Overview

  • The research focused on investigating the impact of different transport conditions on the biological properties of mesenchymal stromal cells (MSCs) derived from the bone marrow of horses. These cells are frequently used in equine treatments.
  • The study tested a range of transport-related parameters including the container used for cell transportation, the medium cells were immersed in, the length of time cells were transported, the temperature during transportation, and the concentration of cells.
  • The research was split into three parts; each examining different factors around transport of stem cells.

Study Methodology

  • The first part of the study compared five different types of transport containers to evaluate their impact on the characteristics of the MSCs.
  • The second part tested seven different transport media, two different temperatures and four different time frames.
  • The third part of the study examined the impact of three different cell concentrations.
  • The cell viability, proliferation potential, and trilineage differentiation capacity were assessed under each test condition.

Results

  • The first part of the study found no significant differences between any of the transport containers when kept at room temperature.
  • The second part of the study found that neither media, temperatures, or time frames had a significant influence on cell viability.
  • The third part of the study demonstrated that different cell concentrations also had no appreciable impact on the measured parameters.
  • However, it was observed that all freezing protocols resulted in unacceptably low viability, particularly with bone marrow supernatant or plasma and DMSO.
  • A trend of decreased chondrogenic differentiation was observed under all transport conditions.

Discussion

  • The results of this study suggest that the transport conditions had minimal influence on the viability, proliferation or trilineage differentiation capabilities of MSCs. This is unexpected as the vulnerability of the cells to changes in their environment is often assumed.
  • The study also highlighted potential issues with freezing protocols and their impact on cell viability. This suggests that variations in freezing or thawing methods could be a factor and warrants further investigation.
  • Additionally, the significance of the decreased chondrogenic differentiation should be explored in the future.

Cite This Article

APA
Espina M, Jülke H, Brehm W, Ribitsch I, Winter K, Delling U. (2016). Evaluation of transport conditions for autologous bone marrow-derived mesenchymal stromal cells for therapeutic application in horses. PeerJ, 4, e1773. https://doi.org/10.7717/peerj.1773

Publication

PeerJ
ISSN: 2167-8359
NlmUniqueID: 101603425
Country: United States
Language: English
Volume: 4
Pages: e1773
PII: e1773

Researcher Affiliations

Espina, Miguel
  • Large Animal Clinic for Surgery, Faculty of Veterinary Medicine, University of Leipzig , Leipzig , Germany.
Jülke, Henriette
  • Translational Centre for Regenerative Medicine (TRM), University of Leipzig , Leipzig , Germany.
Brehm, Walter
  • Large Animal Clinic for Surgery, Faculty of Veterinary Medicine, University of Leipzig , Leipzig , Germany.
Ribitsch, Iris
  • Translational Centre for Regenerative Medicine (TRM), University of Leipzig, Leipzig, Germany; Equine Clinic, University of Veterinary Medicine Vienna, Vienna, Austria.
Winter, Karsten
  • Translational Centre for Regenerative Medicine (TRM), University of Leipzig, Leipzig, Germany; Institute of Anatomy, Faculty of Medicine, University of Leipzig, Leipzig, Germany.
Delling, Uta
  • Large Animal Clinic for Surgery, Faculty of Veterinary Medicine, University of Leipzig , Leipzig , Germany.

Conflict of Interest Statement

The authors declare there are no competing interests.

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Citations

This article has been cited 10 times.
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