Introduction: Autologous mesenchymal stem cell (MSC) injection into naturally-occurring equine tendon injuries\nhas been shown to be safe and efficacious and protocols inform translation of the technique into humans. Efficient\ntransfer of cells from the laboratory into tissue requires well-validated transport and implantation techniques.\nMethods: Cell viability in a range of media was determined over 72 hours and after injection through a 19G,\n21G or 23G needle. Viability, proliferation and apoptosis were analysed using TrypanBlue, alamarBlue�® and\nAnnexinV assays.\nResults: Cell viability was similar in all re-suspension media following 24 hour storage, however cell death was\nmost rapid in bone marrow aspirate, platelet-rich plasma and serum after longer storage. Cryogenic media exhibited\ngreatest viability regardless of storage time. Cell proliferation after 24 and 72 hour storage was similar for all media,\nexcept after 24 hours in serum wherein proliferation was enhanced. MSC tri-lineage differentiation and viability did\nnot significantly change when extruded through 19G, 21G or 23G needles, but 21G and 23G needles significantly\nincreased apoptotic cells compared to 19G and non-injected controls. All gauges induced a decrease in metabolic\nactivity immediately post-injection but cells recovered by 2 hours.\nConclusions: These data indicate storage and injection influence viability and subsequent cell behaviour and\nprovide recommendations for MSC therapy that implantation of cells should occur within 24 hours of recovery\nfrom culture, using larger needle bores.
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