FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Stem Cell Res Ther / Viability of equine mesenchymal stem cells during transport ...
Stem cell research & therapy2014; 5(4); 94; doi: 10.1186/scrt483

Viability of equine mesenchymal stem cells during transport and implantation.

Abstract: Autologous mesenchymal stem cell (MSC) injection into naturally-occurring equine tendon injuries has been shown to be safe and efficacious and protocols inform translation of the technique into humans. Efficient transfer of cells from the laboratory into tissue requires well-validated transport and implantation techniques. Methods: Cell viability in a range of media was determined over 72 hours and after injection through a 19G, 21G or 23G needle. Viability, proliferation and apoptosis were analysed using TrypanBlue, alamarBlue® and AnnexinV assays. Results: Cell viability was similar in all re-suspension media following 24 hour storage, however cell death was most rapid in bone marrow aspirate, platelet-rich plasma and serum after longer storage. Cryogenic media exhibited greatest viability regardless of storage time. Cell proliferation after 24 and 72 hour storage was similar for all media, except after 24 hours in serum wherein proliferation was enhanced. MSC tri-lineage differentiation and viability did not significantly change when extruded through 19G, 21G or 23G needles, but 21G and 23G needles significantly increased apoptotic cells compared to 19G and non-injected controls. All gauges induced a decrease in metabolic activity immediately post-injection but cells recovered by 2 hours. Conclusions: These data indicate storage and injection influence viability and subsequent cell behaviour and provide recommendations for MSC therapy that implantation of cells should occur within 24 hours of recovery from culture, using larger needle bores.
Get Full Text
Publication Date: 2014-08-08 PubMed ID: 25107289PubMed Central: PMC4247703DOI: 10.1186/scrt483Google 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 article discusses the survival rate of equine mesenchymal stem cells during storage, transportation, and injection for the treatment of horse tendon injuries. The findings provide recommendations for maintaining stem cell viability and overall effectiveness of the therapy.

Research Methods

  • The researchers examined stem cell viability in different media over 72 hours and following injection through a variety of needle gauges (19G, 21G and 23G).
  • Cell survival, proliferation, and apoptosis were analyzed using three different assays: TrypanBlue, alamarBlue®, and AnnexinV.

Research Findings

  • Cell viability in the first 24-hours was similar across all resuspension media. However, cells experienced rapid death in bone marrow aspirate, platelet-rich plasma, and serum after longer storage.
  • Cryogenic media maintained the highest cell survival rate irrespective of storage time.
  • Except for serum, cell proliferation was similar for all media after storage for 24 and 72 hours. The proliferation was enhanced in serum after 24 hours.
  • The differentiation and viability of the stem cells did not significantly change when injected through any of the needle gauges (19G, 21G, or 23G). However, more apoptotic cells were observed when the cells were injected through the 21G and 23G needles as compared to the 19G needle and the non-injected controls.
  • All the needle gauges induced a temporary decline in metabolic activity immediately after injection, but the cells managed to recover within two hours.

Conclusions

  • The study findings reveal that storage media and injection methods can significantly influence the viability of the stem cells and their subsequent behavior.
  • The authors recommend that for maintaining the effectiveness of equine mesenchymal stem cell therapy, the cells should be implanted within 24 hours of recovery from culture and larger needle bores should be used for injection.

Cite This Article

APA
Garvican ER, Cree S, Bull L, Smith RK, Dudhia J. (2014). Viability of equine mesenchymal stem cells during transport and implantation. Stem Cell Res Ther, 5(4), 94. https://doi.org/10.1186/scrt483

Publication

Stem cell research & therapy
ISSN: 1757-6512
NlmUniqueID: 101527581
Country: England
Language: English
Volume: 5
Issue: 4
Pages: 94

Researcher Affiliations

Garvican, Elaine R
    Cree, Sandra
      Bull, Lydia
        Smith, Roger Kw
          Dudhia, Jayesh

            MeSH Terms

            • Animals
            • Cell Culture Techniques
            • Cell Differentiation
            • Cell Survival
            • Cell- and Tissue-Based Therapy
            • Cryopreservation
            • Culture Media
            • Horses
            • Mesenchymal Stem Cell Transplantation / instrumentation
            • Mesenchymal Stem Cells / cytology
            • Needles
            • Specimen Handling / methods
            • Tendon Injuries / therapy

            Grant Funding

            • Medical Research Council

            References

            This article includes 27 references
            1. Godwin EE, Young NJ, Dudhia J, Beamish IC, Smith RK. Implantation of bone marrow-derived mesenchymal stem cells demonstrates improved outcome in horses with overstrain injury of the superficial digital flexor tendon.. Equine Vet J 2012;44:25–32.
              doi: 10.1111/j.2042-3306.2011.00363.xpubmed: 21615465google scholar: lookup
            2. Ouyang HW, Goh JC, Thambyah A, Teoh SH, Lee EH. Knitted poly-lactide-co-glycolide scaffold loaded with bone marrow stromal cells in repair and regeneration of rabbit Achilles tendon.. Tissue Eng 2003;9:431–439.
              doi: 10.1089/107632703322066615pubmed: 12857411google scholar: lookup
            3. Hankemeier S, Keus M, Zeichen J, Jagodzinski M, Barkhausen T, Bosch U, Krettek C, Van Griensven M. Modulation of proliferation and differentiation of human bone marrow stromal cells by fibroblast growth factor 2: potential implications for tissue engineering of tendons and ligaments.. Tissue Eng 2005;11:41–49.
              doi: 10.1089/ten.2005.11.41pubmed: 15738660google scholar: lookup
            4. Young RG, Butler DL, Weber W, Caplan AI, Gordon SL, Fink DJ. Use of mesenchymal stem cells in a collagen matrix for Achilles tendon repair.. J Orthop Res 1998;16:406–413.
              doi: 10.1002/jor.1100160403pubmed: 9747780google scholar: lookup
            5. Awad HA, Butler DL, Boivin GP, Smith FN, Malaviya P, Huibregtse B, Caplan AI. Autologous mesenchymal stem cell-mediated repair of tendon.. Tissue Eng 1999;5:267–277.
              doi: 10.1089/ten.1999.5.267pubmed: 10434073google scholar: lookup
            6. Lundborg G, Rank F. Experimental intrinsic healing of flexor tendons based upon synovial fluid nutrition.. J Hand Surg Am 1978;3:21–31.
              doi: 10.1016/S0363-5023(78)80114-2pubmed: 621365google scholar: lookup
            7. Kajikawa Y, Morihara T, Watanabe N, Sakamoto H, Matsuda K, Kobayashi M, Oshima Y, Yoshida A, Kawata M, Kubo T. GFP chimeric models exhibited a biphasic pattern of mesenchymal cell invasion in tendon healing.. J Cell Physiol 2007;210:684–691.
              doi: 10.1002/jcp.20876pubmed: 17154365google scholar: lookup
            8. Lui PP, Ng SW. Cell therapy for the treatment of tendinopathy – a systematic review on the pre-clinical and clinical evidence.. Semin Arthritis Rheum 2013;42:651–666.
              doi: 10.1016/j.semarthrit.2012.10.004pubmed: 23369658google scholar: lookup
            9. Smith RK, Korda M, Blunn GW, Goodship AE. Isolation and implantation of autologous equine mesenchymal stem cells from bone marrow into the superficial digital flexor tendon as a potential novel treatment.. Equine Vet J 2003;35:99–102.
              doi: 10.2746/042516403775467388pubmed: 12553472google scholar: lookup
            10. Walker PA, Jimenez F, Gerber MH, Aroom KR, Shah SK, Harting MT, Gill BS, Savitz SI, Cox CS. Effect of needle diameter and flow rate on rat and human mesenchymal stromal cell characterisation and viability.. Tissue Eng 2010;16:989–997.
              doi: 10.1089/ten.tec.2009.0423pmc: PMC2943405pubmed: 20001789google scholar: lookup
            11. Alves AG, Stewart AA, Dudhia J, Kasashima Y, Goodship AE, Smith RK. Cell-based therapies for tendon and ligament injuries.. Vet Clin North Am Equine Pract 2011;27:315–333.
              doi: 10.1016/j.cveq.2011.06.001pubmed: 21872761google scholar: lookup
            12. Becerra P, Valdés MA, Fiske-Jackson AR, Dudhia J, Neves F, Hartman NG, Smith RKW. The distribution of injected technetium99m-labelled mesenchymal stem cells in horses with naturally-occurring tendinopathy.. J Orthop Res 2013;31:1096–1102.
              doi: 10.1002/jor.22338pubmed: 23508674google scholar: lookup
            13. Frisbie DD, Kisiday JD, Kawcak CE, McIlwraith CW, Werpy NM. Evaluation of adipose derived stromal vascular fraction or bone marrow derived mesenchymal stem cells for treatment of osteoarthritis.. J Orthop Res 2009;27:1675–1680.
              doi: 10.1002/jor.20933pubmed: 19544397google scholar: lookup
            14. Elmore S. Apoptosis: a review of programmed cell death.. Toxicol Pathol 2007;35:495–516.
              doi: 10.1080/01926230701320337pmc: PMC2117903pubmed: 17562483google scholar: lookup
            15. Sharp MK, Mohammad SF. Scaling of hemolysis in needles and catheters.. Ann Biomed Eng 1998;26:788–797.
              doi: 10.1114/1.65pubmed: 9779951google scholar: lookup
            16. Tol M, Akar AR, Durdu S, Ayyildiz E, Ilhan O. Comparison of different needle diameters and flow rates on bone marrow mononuclear stem cell viability: an ex vivo experimental study.. Cytotherapy 2008;10:98–99.
              doi: 10.1080/14653240701762356pubmed: 18202979google scholar: lookup
            17. Kondziolka D, Gobbel GT, Fellows-Mayle W, Chang YF, Uram M. Injection parameters affect cell viability and implant volumes in automated cell delivery for the brain.. Cell Transplant 2011;20:1901–1906.
              doi: 10.3727/096368911X566190pubmed: 21457614google scholar: lookup
            18. Chang R, Nam J, Sun W. Effects of dispensing pressure and nozzle diameter on cell survival from solid freeform fabrication-based direct cell writing.. Tissue Eng Part A 2008;14:41–49.
              doi: 10.1089/ten.a.2007.0004pubmed: 18333803google scholar: lookup
            19. Li M, Tian X, Schreyer DJ, Chen X. Effect of needle geometry on flow rate and cell damage in the dispensing-based biofabrication process.. Biotechnol Prog 2011;27:1777–1784.
              doi: 10.1002/btpr.679pubmed: 22238771google scholar: lookup
            20. Dunkel B, Rickards KJ, Page CP, Cunningham FM. Platelet activation in ponies with airway inflammation.. Equine Vet J 2007;39:557–561.
              doi: 10.2746/042516407X217885pubmed: 18065316google scholar: lookup
            21. Strober W. Trypan blue exclusion test of cell viability.. Curr Protoc Immunol 2001;21:3B:A.3B.1–A.3B.2.
              pubmed: 18432654
            22. Smith RK, Werling NJ, Dakin SG, Alam R, Goodship AE, Dudhia J. Beneficial effects of autologous bone marrow-derived mesenchymal stem cells in naturally occurring tendinopathy.. PLoS One 2013;8:e75697.
              doi: 10.1371/journal.pone.0075697pmc: PMC3783421pubmed: 24086616google scholar: lookup
            23. Pamphilon DH, Selogie E, Szczepiorkowski ZM. Transportation of cellular therapy products: report of a survey by the cellular therapies team of the Biomedical Excellence for Safer Transfusion (BEST) collaborative.. Vox Sang 2010;99:168–173.
              doi: 10.1111/j.1423-0410.2010.01329.xpubmed: 20230598google scholar: lookup
            24. Sohn HS, Heo JS, Kim HS, Choi Y, Kim HO. Duration of in vitro storage affects the key stem cell features of human bone marrow-derived mesenchymal stromal cells for clinical transplantation.. Cytotherapy 2013;15:460–466.
              doi: 10.1016/j.jcyt.2012.10.015pubmed: 23318345google scholar: lookup
            25. Muraki K, Hirose M, Kotobuki N, Kato Y, Machida H, Takakura Y. Assessment of viability and osteogenic ability of human mesenchymal stem cells after being stored in suspension for clinical transplantation.. Tissue Eng 2006;12:1711–1719.
              doi: 10.1089/ten.2006.12.1711pubmed: 16846365google scholar: lookup
            26. Pal R, Hanwate M, Totey SM. Effect of holding time, temperature and different parenteral solutions on viability and functionality of adult bone marrow-derived mesenchymal stem cells before transplantation.. J Tissue Eng Regen Med 2008;7:436–444.
              doi: 10.1002/term.109pubmed: 18720444google scholar: lookup
            27. Zhang H, Kay A, Forsyth NR, Liu KK, El Haj AJ. Gene expression of single human mesenchymal stem cell in response to fluid shear.. J Tissue Eng 2012;3:2041731412451988.
              doi: 10.1177/2041731412451988pmc: PMC3394398pubmed: 22798982google scholar: lookup

            Citations

            This article has been cited 30 times.
            1. Srinivasan S, Ghone NV. Modulating Proliferation, Migration and Differentiation of Mesenchymal Stem Cells Using Interleukins. Curr Stem Cell Res Ther 2025;20(5):546-564.
              doi: 10.2174/011574888X313750240524115446pubmed: 40525425google scholar: lookup
            2. Aabling RR, Rusan M, Møller AMJ, Munk-Pedersen N, Holm C, Elmengaard B, Pedersen M, Møller BK. A Narrative Review on Manufacturing Methods Employed in the Production of Mesenchymal Stromal Cells for Knee Osteoarthritis Therapy. Biomedicines 2025 Feb 18;13(2).
              doi: 10.3390/biomedicines13020509pubmed: 40002922google scholar: lookup
            3. Quam VG, Belacic ZA, Long S, Rice HC, Dhar MS, Durgam S. Equine bone marrow MSC-derived extracellular vesicles mitigate the inflammatory effects of interleukin-1β on navicular tissues in vitro. Equine Vet J 2025 Jan;57(1):232-242.
              doi: 10.1111/evj.14090pubmed: 38587145google scholar: lookup
            4. Andreoli V, Berni P, Conti V, Ramoni R, Basini G, Grolli S. Mesenchymal Stromal Cells Derived from Canine Adipose Tissue: Evaluation of the Effect of Different Shipping Vehicles Used for Clinical Administration. Int J Mol Sci 2024 Mar 18;25(6).
              doi: 10.3390/ijms25063426pubmed: 38542399google scholar: lookup
            5. Taufiq H, Shaik Fakiruddin K, Muzaffar U, Lim MN, Rusli S, Kamaluddin NR, Esa E, Abdullah S. Systematic review and meta-analysis of mesenchymal stromal/stem cells as strategical means for the treatment of COVID-19. Ther Adv Respir Dis 2023 Jan-Dec;17:17534666231158276.
              doi: 10.1177/17534666231158276pubmed: 37128999google scholar: lookup
            6. Iacono E, Merlo B. Stem Cells in Domestic Animals: Applications in Health and Production. Animals (Basel) 2022 Oct 13;12(20).
              doi: 10.3390/ani12202753pubmed: 36290139google scholar: lookup
            7. Iacono E, Lanci A, Gugole P, Merlo B. Shipping Temperature, Time and Media Effects on Equine Wharton's Jelly and Adipose Tissue Derived Mesenchymal Stromal Cells Characteristics. Animals (Basel) 2022 Aug 3;12(15).
              doi: 10.3390/ani12151967pubmed: 35953956google scholar: lookup
            8. Ivanovska A, Wang M, Arshaghi TE, Shaw G, Alves J, Byrne A, Butterworth S, Chandler R, Cuddy L, Dunne J, Guerin S, Harry R, McAlindan A, Mullins RA, Barry F. Manufacturing Mesenchymal Stromal Cells for the Treatment of Osteoarthritis in Canine Patients: Challenges and Recommendations. Front Vet Sci 2022;9:897150.
              doi: 10.3389/fvets.2022.897150pubmed: 35754551google scholar: lookup
            9. De Paula JC, Doello K, Mesas C, Kapravelou G, Cornet-Gómez A, Orantes FJ, Martínez R, Linares F, Prados JC, Porres JM, Osuna A, de Pablos LM. Exploring Honeybee Abdominal Anatomy through Micro-CT and Novel Multi-Staining Approaches. Insects 2022 Jun 18;13(6).
              doi: 10.3390/insects13060556pubmed: 35735893google scholar: lookup
            10. Bedar M, Jerez S, Pulos N, van Wijnen AJ, Shin AY. Dynamic seeding versus microinjection of mesenchymal stem cells for acellular nerve allograft: an in vitro comparison. J Plast Reconstr Aesthet Surg 2022 Aug;75(8):2821-2830.
              doi: 10.1016/j.bjps.2022.04.017pubmed: 35570113google scholar: lookup
            11. Sultana T, Dayem AA, Lee SB, Cho SG, Lee JI. Effects of carrier solutions on the viability and efficacy of canine adipose-derived mesenchymal stem cells. BMC Vet Res 2022 Jan 7;18(1):26.
              doi: 10.1186/s12917-021-03120-4pubmed: 34996443google scholar: lookup
            12. Wasai S, Toyoda E, Takahashi T, Maehara M, Okada E, Uchiyama R, Akamatsu T, Watanabe M, Sato M. Development of Injectable Polydactyly-Derived Chondrocyte Sheets. Int J Mol Sci 2021 Mar 21;22(6).
              doi: 10.3390/ijms22063198pubmed: 33801144google scholar: lookup
            13. Wang H, Zhu D, Paul A, Cai L, Enejder A, Yang F, Heilshorn SC. Covalently adaptable elastin-like protein - hyaluronic acid (ELP - HA) hybrid hydrogels with secondary thermoresponsive crosslinking for injectable stem cell delivery. Adv Funct Mater 2017 Jul 26;27(28).
              doi: 10.1002/adfm.201605609pubmed: 33041740google scholar: lookup
            14. Mocchi M, Dotti S, Bue MD, Villa R, Bari E, Perteghella S, Torre ML, Grolli S. Veterinary Regenerative Medicine for Musculoskeletal Disorders: Can Mesenchymal Stem/Stromal Cells and Their Secretome Be the New Frontier?. Cells 2020 Jun 11;9(6).
              doi: 10.3390/cells9061453pubmed: 32545382google scholar: lookup
            15. Mathot F, Rbia N, Thaler R, Bishop AT, van Wijnen AJ, Shin AY. Introducing human adipose-derived mesenchymal stem cells to Avance(Ⓡ) nerve grafts and NeuraGen(Ⓡ) nerve guides. J Plast Reconstr Aesthet Surg 2020 Aug;73(8):1473-1481.
              doi: 10.1016/j.bjps.2020.03.012pubmed: 32418840google scholar: lookup
            16. Kamm JL, Parlane NA, Riley CB, Gee EK, Dittmer KE, McIlwraith CW. Blood type and breed-associated differences in cell marker expression on equine bone marrow-derived mesenchymal stem cells including major histocompatibility complex class II antigen expression. PLoS One 2019;14(11):e0225161.
              doi: 10.1371/journal.pone.0225161pubmed: 31747418google scholar: lookup
            17. Shojaee A, Parham A. Strategies of tenogenic differentiation of equine stem cells for tendon repair: current status and challenges. Stem Cell Res Ther 2019 Jun 18;10(1):181.
              doi: 10.1186/s13287-019-1291-0pubmed: 31215490google scholar: lookup
            18. Mathot F, Rbia N, Bishop AT, Hovius SER, Van Wijnen AJ, Shin AY. Adhesion, distribution, and migration of differentiated and undifferentiated mesenchymal stem cells (MSCs) seeded on nerve allografts. J Plast Reconstr Aesthet Surg 2020 Jan;73(1):81-89.
              doi: 10.1016/j.bjps.2019.05.030pubmed: 31202698google scholar: lookup
            19. Mathot F, Shin AY, Van Wijnen AJ. Targeted stimulation of MSCs in peripheral nerve repair. Gene 2019 Aug 20;710:17-23.
              doi: 10.1016/j.gene.2019.02.078pubmed: 30849542google scholar: lookup
            20. Bogers SH. Cell-Based Therapies for Joint Disease in Veterinary Medicine: What We Have Learned and What We Need to Know. Front Vet Sci 2018;5:70.
              doi: 10.3389/fvets.2018.00070pubmed: 29713634google scholar: lookup
            21. Santos VH, Pfeifer JPH, de Souza JB, Milani BHG, de Oliveira RA, Assis MG, Deffune E, Moroz A, Alves ALG. Culture of mesenchymal stem cells derived from equine synovial membrane in alginate hydrogel microcapsules. BMC Vet Res 2018 Mar 27;14(1):114.
              doi: 10.1186/s12917-018-1425-0pubmed: 29587733google scholar: lookup
            22. Zhang F, Ren H, Shao X, Zhuang C, Chen Y, Qi N. Preservation media, durations and cell concentrations of short-term storage affect key features of human adipose-derived mesenchymal stem cells for therapeutic application. PeerJ 2017;5:e3301.
              doi: 10.7717/peerj.3301pubmed: 28533959google scholar: lookup
            23. Lang HM, Schnabel LV, Cassano JM, Fortier LA. Effect of needle diameter on the viability of equine bone marrow derived mesenchymal stem cells. Vet Surg 2017 Jul;46(5):731-737.
              doi: 10.1111/vsu.12639pubmed: 28328147google scholar: lookup
            24. Garvican ER, Cree S, Bull L, Smith RK, Dudhia J. Erratum to: Viability of equine mesenchymal stem cells during transport and implantation. Stem Cell Res Ther 2016 Nov 9;7(1):161.
              doi: 10.1186/s13287-016-0423-zpubmed: 27829446google scholar: lookup
            25. 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
            26. Markoski MM. Advances in the Use of Stem Cells in Veterinary Medicine: From Basic Research to Clinical Practice. Scientifica (Cairo) 2016;2016:4516920.
              doi: 10.1155/2016/4516920pubmed: 27379197google scholar: lookup
            27. Espina M, Jülke H, Brehm W, Ribitsch I, Winter K, Delling U. Evaluation of transport conditions for autologous bone marrow-derived mesenchymal stromal cells for therapeutic application in horses. PeerJ 2016;4:e1773.
              doi: 10.7717/peerj.1773pubmed: 27019778google scholar: lookup
            28. 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
            29. Onishi K, Jones DL, Riester SM, Lewallen EA, Lewallen DG, Sellon JL, Dietz AB, Qu W, van Wijnen AJ, Smith J. Human Adipose-Derived Mesenchymal Stromal/Stem Cells Remain Viable and Metabolically Active Following Needle Passage. PM R 2016 Sep;8(9):844-54.
              doi: 10.1016/j.pmrj.2016.01.010pubmed: 26826615google scholar: lookup
            30. Mitchell A, Rivas KA, Smith R 3rd, Watts AE. Cryopreservation of equine mesenchymal stem cells in 95% autologous serum and 5% DMSO does not alter post-thaw growth or morphology in vitro compared to fetal bovine serum or allogeneic serum at 20 or 95% and DMSO at 10 or 5. Stem Cell Res Ther 2015 Nov 26;6:231.
              doi: 10.1186/s13287-015-0230-ypubmed: 26611913google scholar: lookup
            Subscribe to our newsletter
            Join 99,000 horse owners like you who receive our email updates on horse health and nutrition.