title: Viability of equine mesenchymal stem cells during transport and implantation.
abstract: BACKGROUND
  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.
authors.forename: Elaine R
authors.initials: ER
authors.lastname: Garvican
authors.forename: Sandra
authors.initials: S
authors.lastname: Cree
authors.forename: Lydia
authors.initials: L
authors.lastname: Bull
authors.forename: Roger Kw
authors.initials: RK
authors.lastname: Smith
authors.forename: Jayesh
authors.initials: J
authors.lastname: Dudhia
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002470:Cell Survival; D064987:Cell- and Tissue-Based Therapy; D015925:Cryopreservation; D003470:Culture Media; D006736:Horses; D045164:Mesenchymal Stem Cell Transplantation / Q000295:instrumentation; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D009339:Needles; D013048:Specimen Handling / Q000379:methods; D013708:Tendon Injuries / Q000628:therapy*
chemical_list: D003470:Culture Media
references.citation: Cytotherapy. 2008;10(1):98-9
references.pmid: 18202979
references.citation: Tissue Eng. 2003 Jun;9(3):431-9
references.pmid: 12857411
references.citation: Semin Arthritis Rheum. 2013 Jun;42(6):651-66
references.pmid: 23369658
references.citation: PLoS One. 2013 Sep 25;8(9):e75697
references.pmid: 24086616
references.citation: J Cell Physiol. 2007 Mar;210(3):684-91
references.pmid: 17154365
references.citation: Tissue Eng. 2006 Jun;12(6):1711-9
references.pmid: 16846365
references.citation: Vet Clin North Am Equine Pract. 2011 Aug;27(2):315-33
references.pmid: 21872761
references.citation: J Tissue Eng Regen Med. 2008 Oct;2(7):436-44
references.pmid: 18720444
references.citation: Vox Sang. 2010 Aug 1;99(2):168-73
references.pmid: 20230598
references.citation: J Orthop Res. 2009 Dec;27(12):1675-80
references.pmid: 19544397
references.citation: Cytotherapy. 2013 Apr;15(4):460-6
references.pmid: 23318345
references.citation: Equine Vet J. 2003 Jan;35(1):99-102
references.pmid: 12553472
references.citation: Cell Transplant. 2011;20(11-12):1901-6
references.pmid: 21457614
references.citation: Toxicol Pathol. 2007 Jun;35(4):495-516
references.pmid: 17562483
references.citation: Tissue Eng. 1999 Jun;5(3):267-77
references.pmid: 10434073
references.citation: Equine Vet J. 2007 Nov;39(6):557-61
references.pmid: 18065316
references.citation: Curr Protoc Immunol. 2001 May;Appendix 3:Appendix 3B
references.pmid: 18432654
references.citation: Tissue Eng Part C Methods. 2010 Oct;16(5):989-97
references.pmid: 20001789
references.citation: Ann Biomed Eng. 1998 Sep-Oct;26(5):788-97
references.pmid: 9779951
references.citation: J Orthop Res. 2013 Jul;31(7):1096-102
references.pmid: 23508674
references.citation: J Hand Surg Am. 1978 Jan;3(1):21-31
references.pmid: 621365
references.citation: J Orthop Res. 1998 Jul;16(4):406-13
references.pmid: 9747780
references.citation: Equine Vet J. 2012 Jan;44(1):25-32
references.pmid: 21615465
references.citation: Biotechnol Prog. 2011 Nov-Dec;27(6):1777-84
references.pmid: 22238771
references.citation: Tissue Eng Part A. 2008 Jan;14(1):41-8
references.pmid: 18333803
references.citation: Tissue Eng. 2005 Jan-Feb;11(1-2):41-9
references.pmid: 15738660
references.citation: J Tissue Eng. 2012;3(1):2041731412451988
references.pmid: 22798982