Infusion of Mesenchymal Stem Cells Protects Lung Transplants from Cold Ischemia-Reperfusion Injury in Mice
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Cold ischemia-reperfusion injury (IRI) is a major cause of graft failure in lung transplantation. Despite therapeutic benefits of mesenchymal stem cells (MSCs) in attenuating acute lung injury, their protection of lung transplants from cold IRI remains elusive. The present study was to test the efficacy of MSCs in the prevention of cold IRI using a novel murine model of orthotopic lung transplantation.
Donor lungs from C57BL/6 mice were exposed to 6 h of cold ischemia before transplanted to syngeneic recipients. MSCs were isolated from the bone marrows of C57BL/6 mice for recipient treatment. Gas exchange was determined by the measurement of blood oxygenation, and lung injury and inflammation were assessed by histological analyses.
Intravenously delivered MSC migration/trafficking to the lung grafts occurred within 4-hours post-transplantation. As compared to untreated controls, the graft arterial blood oxygenation (PaO2/FiO2) capacity was significantly improved in MSC-treated recipients as early as 4 h post-reperfusion and such improvement continued over time. By 72 h, oxygenation reached normal level that was not seen in controls. MSCs treatment conferred significant protection of the grafts from cold IRI and cell apoptosis, which is correlated with less cellular infiltration, a decrease in proinflammatory cytokines (TNF-α, IL-6) and toll-like receptor 4, and an increase in anti-inflammatory TSG-6 generation.
MSCs provide significant protection against cold IRI in lung transplants, and thus may be a promising strategy to improve outcomes after lung transplantation.
KeywordsMesenchymal stem cells Orthotopic lung transplantation Cold ischemia-reperfusion injury Mice
Analysis of variance
Enhanced green fluorescence protein
Hematoxylin & eosin
Mesenchymal stem cells
Reactive oxygen species
Standard error of mean
Tumor necrosis factor
Tumor necrosis factor-α stimulated gene/protein
Terminal deoxynucleotyl transferase dUTP nick end labeling
We are grateful to Dr. Bertha Garcia at Western University, London, Canada for her critical review of the histological slides, and to Drs. Shuhua Luo and Weihua Liu for their technical assistance. This work was supported by Grant to H.W. from National Natural Science Foundation of China (No. 81273257), and National High Technology Research and Development Program 863 (2012AA021003). C.D. was supported by grants from the Kidney Foundation of Canada and the Canadian Institutes of Health Research.
Conflict of interest
The authors have no conflicts of interest to disclose.
- 5.Yip HK, Chang YC, Wallace CG, Chang LT, Tsai TH, Chen YL, Chang HW, Leu S, Zhen YY, Tsai CY, Yeh KH, Sun CK, Yen CH (2013) Melatonin treatment improves adipose-derived mesenchymal stem cell therapy for acute lung ischemia-reperfusion injury. J Pineal Res 54(2):207–221. doi: 10.1111/jpi.12020 PubMedCrossRefGoogle Scholar
- 8.Svobodova E, Krulova M, Zajicova A, Pokorna K, Prochazkova J, Trosan P, Holan V (2012) The role of mouse mesenchymal stem cells in differentiation of naive T-cells into anti-inflammatory regulatory T-cell or proinflammatory helper T-cell 17 population. Stem Cells Dev 21(6):901–910. doi: 10.1089/scd.2011.0157 PubMedCentralPubMedCrossRefGoogle Scholar
- 12.Assis AC, Carvalho JL, Jacoby BA, Ferreira RL, Castanheira P, Diniz SO, Cardoso VN, Goes AM, Ferreira AJ (2010) Time-dependent migration of systemically delivered bone marrow mesenchymal stem cells to the infarcted heart. Cell Transplant 19(2):219–230. doi: 10.3727/096368909X479677 PubMedCrossRefGoogle Scholar
- 13.Lee RH, Pulin AA, Seo MJ, Kota DJ, Ylostalo J, Larson BL, Semprun-Prieto L, Delafontaine P, Prockop DJ (2009) Intravenous hMSCs improve myocardial infarction in mice because cells embolized in lung are activated to secrete the anti-inflammatory protein TSG-6. Cell Stem Cell 5(1):54–63. doi: 10.1016/j.stem.2009.05.003 PubMedCentralPubMedCrossRefGoogle Scholar
- 14.Barbash IM, Chouraqui P, Baron J, Feinberg MS, Etzion S, Tessone A, Miller L, Guetta E, Zipori D, Kedes LH, Kloner RA, Leor J (2003) Systemic delivery of bone marrow-derived mesenchymal stem cells to the infarcted myocardium: feasibility, cell migration, and body distribution. Circulation 108(7):863–868. doi: 10.1161/01.CIR.0000084828.50310.6A PubMedCrossRefGoogle Scholar
- 15.Fischer UM, Harting MT, Jimenez F, Monzon-Posadas WO, Xue H, Savitz SI, Laine GA, Cox CS Jr (2009) Pulmonary passage is a major obstacle for intravenous stem cell delivery: the pulmonary first-pass effect. Stem Cells Dev 18(5):683–692. doi: 10.1089/scd.2008.0253 PubMedCentralPubMedCrossRefGoogle Scholar
- 16.Ge W, Jiang J, Baroja ML, Arp J, Zassoko R, Liu W, Bartholomew A, Garcia B, Wang H (2009) Infusion of mesenchymal stem cells and rapamycin synergize to attenuate alloimmune responses and promote cardiac allograft tolerance. Am J Transplant 9(8):1760–1772. doi: 10.1111/j.1600-6143.2009.02721.x PubMedCrossRefGoogle Scholar
- 18.Chen S, Chen L, Wu X, Lin J, Fang J, Chen X, Wei S, Xu J, Gao Q, Kang M (2012) Ischemia postconditioning and mesenchymal stem cells engraftment synergistically attenuate ischemia reperfusion-induced lung injury in rats. J Surg Res 178(1):81–91. doi: 10.1016/j.jss.2012.01.039 PubMedCrossRefGoogle Scholar
- 20.Zanotti G, Casiraghi M, Abano JB, Tatreau JR, Sevala M, Berlin H, Smyth S, Funkhouser WK, Burridge K, Randell SH, Egan TM (2009) Novel critical role of Toll-like receptor 4 in lung ischemia-reperfusion injury and edema. Am J Physiol Lung Cell Mol Physiol 297(1):L52–63. doi: 10.1152/ajplung.90406.2008 PubMedCentralPubMedCrossRefGoogle Scholar
- 21.Getting SJ, Mahoney DJ, Cao T, Rugg MS, Fries E, Milner CM, Perretti M, Day AJ (2002) The link module from human TSG-6 inhibits neutrophil migration in a hyaluronan- and inter-alpha -inhibitor-independent manner. J Biol Chem 277(52):51068–51076. doi: 10.1074/jbc.M205121200 PubMedCrossRefGoogle Scholar
- 22.Choi H, Lee RH, Bazhanov N, Oh JY, Prockop DJ (2011) Anti-inflammatory protein TSG-6 secreted by activated MSCs attenuates zymosan-induced mouse peritonitis by decreasing TLR2/NF-kappaB signaling in resident macrophages. Blood 118(2):330–338. doi: 10.1182/blood-2010-12-327353 PubMedCentralPubMedCrossRefGoogle Scholar
- 23.Sun CK, Yen CH, Lin YC, Tsai TH, Chang LT, Kao YH, Chua S, Fu M, Ko SF, Leu S, Yip HK (2011) Autologous transplantation of adipose-derived mesenchymal stem cells markedly reduced acute ischemia-reperfusion lung injury in a rodent model. J Transl Med 9:118. doi: 10.1186/1479-5876-9-118 PubMedCentralPubMedCrossRefGoogle Scholar
- 25.Okazaki M, Krupnick AS, Kornfeld CG, Lai JM, Ritter JH, Richardson SB, Huang HJ, Das NA, Patterson GA, Gelman AE, Kreisel D (2007) A mouse model of orthotopic vascularized aerated lung transplantation. Am J Transplant 7(6):1672–1679. doi: 10.1111/j.1600-6143.2007.01819.x PubMedCrossRefGoogle Scholar
- 26.Krupnick AS, Lin X, Li W, Okazaki M, Lai J, Sugimoto S, Richardson SB, Kornfeld CG, Garbow JR, Patterson GA, Gelman AE, Kreisel D (2009) Orthotopic mouse lung transplantation as experimental methodology to study transplant and tumor biology. Nat Protoc 4(1):86–93. doi: 10.1038/nprot.2008.218 PubMedCrossRefGoogle Scholar
- 30.Kaczorowski DJ, Nakao A, Mollen KP, Vallabhaneni R, Sugimoto R, Kohmoto J, Tobita K, Zuckerbraun BS, McCurry KR, Murase N, Billiar TR (2007) Toll-like receptor 4 mediates the early inflammatory response after cold ischemia/reperfusion. Transplantation 84(10):1279–1287. doi: 10.1097/01.tp.0000287597.87571.17 PubMedCrossRefGoogle Scholar
- 33.Kinnaird T, Stabile E, Burnett MS, Lee CW, Barr S, Fuchs S, Epstein SE (2004) Marrow-derived stromal cells express genes encoding a broad spectrum of arteriogenic cytokines and promote in vitro and in vivo arteriogenesis through paracrine mechanisms. Circ Res 94(5):678–685. doi: 10.1161/01.RES.0000118601.37875.AC PubMedCrossRefGoogle Scholar