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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.
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Publication Date: 2014-08-08 PubMed ID: 25107289PubMed Central: PMC4247703DOI: 10.1186/scrt483Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

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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
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