Name: pre-conditioning the airways of mice with bleomycin increases the efficiency of orthotopic lung cancer cell engraftment
Uploaded: 2018-06-28T12:30:00
Description: Watch this Scientific Journal Video about Pre-Conditioning the Airways of Mice with Bleomycin Increases the Efficiency of Orthotopic Lung Cancer Cell Engraftment at JoVE.com

Este estudio fue financiado por becas del Instituto Nacional del cáncer (R01CA166376 y R01CA191489 a D.X. Nguyen) y el Departamento de defensa (W81XWH-16-1-0227 a D.X. Nguyen).

Todos los experimentos se llevaron a cabo según los protocolos aprobados por el cuidado institucional del Animal y el Comité uso (IACUC) en Universidad de Yale.
1. configuración / preparación de los reactivos.
<ol><li>
Bleomicina PRECAUCIÓN: Basado en el mundialmente armonizado sistema (SAM) de clasificación y etiquetado de productos químicos, bleomicina está clasificada como un riesgo para la salud GHS08.<ol>
<li>Preparación de bleomicina en una campana química...

<ol>
	<li>Siegel, R. L., Miller, K. D., Jemal, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer+statistics,+2015.">Cancer statistics, 2015.</a> CA-Cancer J Clin. 65, 5-29 (2015).</li><li>Gaspar, L. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brain+metastases+in+lung+cancer.">Brain metastases in lung cancer.</a> Expert Rev Anticanc. 4, 259-270 (2004).</li><li>Hess, K. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metastatic+patterns+in+adenocarcinoma.">Metastatic patterns in adenocarcinoma.</a> Cancer. 106, 1624-1633 (2006).</li><li>Hoffman, P. C., Mauer, A. M., Vokes, E. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lung+cancer.">Lung cancer.</a> Lancet. 355, 479-485 (2000).</li><li>Travis, W. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathology+of+lung+cancer.">Pathology of lung cancer.</a> Clin Chest Med. 23 (1), 65-81 (2002).</li><li>Chen, Z., Fillmore, C. M., Hammerman, P. S., Kim, C. F., Wong, K. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-small-cell+lung+cancers:+a+heterogeneous+set+of+diseases.">Non-small-cell lung cancers: a heterogeneous set of diseases.</a> Nat Rev Cancer. 14, 535-546 (2014).</li><li>Kim, C. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+of+human+non-small-cell+lung+cancer:+raising+the+bar.">Mouse models of human non-small-cell lung cancer: raising the bar.</a> Cold Spring Harb Sym. 70, 241-250 (2005).</li><li>Meuwissen, R., Berns, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+for+human+lung+cancer.">Mouse models for human lung cancer.</a> Gene Dev. 19, 643-664 (2005).</li><li>Junttila, M. R., de Sauvage, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+tumour+micro-environment+heterogeneity+on+therapeutic+response.">Influence of tumour micro-environment heterogeneity on therapeutic response.</a> Nature. 501, 346-354 (2013).</li><li>Nguyen, L. V., Vanner, R., Dirks, P., Eaves, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer+stem+cells:+an+evolving+concept.">Cancer stem cells: an evolving concept.</a> Nat Rev Cancer. 12, 133-143 (2012).</li><li>Minchinton, A. I., Tannock, I. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drug+penetration+in+solid+tumours.">Drug penetration in solid tumours.</a> Nat Rev Cancer. 6, 583-592 (2006).</li><li>Leblond, A. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developing+cell+therapy+techniques+for+respiratory+disease:+intratracheal+delivery+of+genetically+engineered+stem+cells+in+a+murine+model+of+airway+injury.">Developing cell therapy techniques for respiratory disease: intratracheal delivery of genetically engineered stem cells in a murine model of airway injury.</a> Hum Gene Ther. 20, 1329-1343 (2009).</li><li>Vaughan, A. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lineage-negative+progenitors+mobilize+to+regenerate+lung+epithelium+after+major+injury.">Lineage-negative progenitors mobilize to regenerate lung epithelium after major injury.</a> Nature. 517, 621-625 (2015).</li><li>Zuo, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=p63(+)Krt5(+)+distal+airway+stem+cells+are+essential+for+lung+regeneration.">p63(+)Krt5(+) distal airway stem cells are essential for lung regeneration.</a> Nature. 517, 616-620 (2015).</li><li>Mahoney, J. E., Kim, C. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tracing+the+potential+of+lung+progenitors.">Tracing the potential of lung progenitors.</a> Nat Biotechnol. 33, 152-154 (2015).</li><li>Wang, D., Morales, J. E., Calame, D. G., Alcorn, J. L., Wetsel, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transplantation+of+human+embryonic+stem+cell-derived+alveolar+epithelial+type+II+cells+abrogates+acute+lung+injury+in+mice.">Transplantation of human embryonic stem cell-derived alveolar epithelial type II cells abrogates acute lung injury in mice.</a> Mol Ther. 18, 625-634 (2010).</li><li>Ortiz, L. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mesenchymal+stem+cell+engraftment+in+lung+is+enhanced+in+response+to+bleomycin+exposure+and+ameliorates+its+fibrotic+effects.">Mesenchymal stem cell engraftment in lung is enhanced in response to bleomycin exposure and ameliorates its fibrotic effects.</a> Proc Natl Acad Sci USA. 100, 8407-8411 (2003).</li><li>Suzuki, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prognostic+significance+of+the+size+of+central+fibrosis+in+peripheral+adenocarcinoma+of+the+lung.">Prognostic significance of the size of central fibrosis in peripheral adenocarcinoma of the lung.</a> Ann Thorac Surg. 69, 893-897 (2000).</li><li>Cancer Genome Atlas Research, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comprehensive+molecular+profiling+of+lung+adenocarcinoma.">Comprehensive molecular profiling of lung adenocarcinoma.</a> Nature. 511, 543-550 (2014).</li><li>Scotton, C. J., Chambers, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bleomycin+revisited:+towards+a+more+representative+model+of+IPF?.">Bleomycin revisited: towards a more representative model of IPF?.</a> Am J Physiol-Lung C. 299, L439-L441 (2010).</li><li>Hay, J., Shahzeidi, S., Laurent, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+bleomycin-induced+lung+damage.">Mechanisms of bleomycin-induced lung damage.</a> Arch Toxicol. 65, 81-94 (1991).</li><li>Vaccaro, C. A., Brody, J. S., Snider, G. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alveolar+wall+basement+membranes+in+bleomycin-induced+pulmonary+fibrosis.">Alveolar wall basement membranes in bleomycin-induced pulmonary fibrosis.</a> Am Rev Respir Dis. 132, 905-912 (1985).</li><li>Venkatesan, N., Ebihara, T., Roughley, P. J., Ludwig, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alterations+in+large+and+small+proteoglycans+in+bleomycin-induced+pulmonary+fibrosis+in+rats.">Alterations in large and small proteoglycans in bleomycin-induced pulmonary fibrosis in rats.</a> Am J Resp Crit Care. 161, 2066-2073 (2000).</li><li>Moore, B. B., Hogaboam, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+models+of+pulmonary+fibrosis.">Murine models of pulmonary fibrosis.</a> Am J Physiol-Lung C. 294, L152-L160 (2008).</li><li>Izbicki, G., Segel, M. J., Christensen, T. G., Conner, M. W., Breuer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time+course+of+bleomycin-induced+lung+fibrosis.">Time course of bleomycin-induced lung fibrosis.</a> Int J Exp Pathol. 83, 111-119 (2002).</li><li>Schrier, D. J., Phan, S. H., McGarry, B. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+the+nude+(nu/nu)+mutation+on+bleomycin-induced+pulmonary+fibrosis.+A+biochemical+evaluation.">The effects of the nude (nu/nu) mutation on bleomycin-induced pulmonary fibrosis. A biochemical evaluation.</a> Am Rev Respir Dis. 127, 614-617 (1983).</li><li>Ponomarev, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+triple-modality+reporter+gene+for+whole-body+fluorescent,+bioluminescent,+and+nuclear+noninvasive+imaging.">A novel triple-modality reporter gene for whole-body fluorescent, bioluminescent, and nuclear noninvasive imaging.</a> Eur J Nucl Med Mol I. 31, 740-751 (2004).</li><li>Morten, B. C., Scott, R. J., Avery-Kiejda, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+Three+Different+Methods+for+Determining+Cell+Proliferation+in+Breast+Cancer+Cell.">Comparison of Three Different Methods for Determining Cell Proliferation in Breast Cancer Cell.</a> J. Vis. Exp. , (2016).</li><li>Tseng, J. C., Kung, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+bioluminescence+imaging+of+mouse+tumor+models.">Quantitative bioluminescence imaging of mouse tumor models.</a> Cold Spring Harbor protocols. , (2015).</li><li>Byrne, F. L., McCarroll, J. A., Kavallaris, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analyses+of+Tumor+Burden+In+Vivo+and+Metastasis+Ex+Vivo+Using+Luciferase-Expressing+Cancer+Cells+in+an+Orthotopic+Mouse+Model+of+Neuroblastoma.">Analyses of Tumor Burden In Vivo and Metastasis Ex Vivo Using Luciferase-Expressing Cancer Cells in an Orthotopic Mouse Model of Neuroblastoma.</a> Methods Mol Biol. 1372, 61-77 (2016).</li><li>Parkinson, C. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diagnostic+necropsy+and+selected+tissue+and+sample+collection+in+rats+and+mice.">Diagnostic necropsy and selected tissue and sample collection in rats and mice.</a> J. Vis. Exp. , (2011).</li><li>Tammela, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Wnt-producing+niche+drives+proliferative+potential+and+progression+in+lung+adenocarcinoma.">A Wnt-producing niche drives proliferative potential and progression in lung adenocarcinoma.</a> Nature. 545, 355-359 (2017).</li><li>DuPage, M., Dooley, A. L., Jacks, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conditional+mouse+lung+cancer+models+using+adenoviral+or+lentiviral+delivery+of+Cre+recombinase.">Conditional mouse lung cancer models using adenoviral or lentiviral delivery of Cre recombinase.</a> Nat Protoc. 4, 1064-1072 (2009).</li><li>Byrne, A. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interrogating+open+issues+in+cancer+precision+medicine+with+patient-derived+xenografts.">Interrogating open issues in cancer precision medicine with patient-derived xenografts.</a> Nat Rev Cancer. 17, 254-268 (2017).</li><li>Nguyen, D. X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=WNT/TCF+signaling+through+LEF1+and+HOXB9+mediates+lung+adenocarcinoma+metastasis.">WNT/TCF signaling through LEF1 and HOXB9 mediates lung adenocarcinoma metastasis.</a> Cell. 138, 51-62 (2009).</li><li>Raghu, G., Nyberg, F., Morgan, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+epidemiology+of+interstitial+lung+disease+and+its+association+with+lung+cancer.">The epidemiology of interstitial lung disease and its association with lung cancer.</a> Brit J Cancer. 91, S3-S10 (2004).</li><li>Hubbard, R., Venn, A., Lewis, S., Britton, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lung+cancer+and+cryptogenic+fibrosing+alveolitis.+A+population-based+cohort+study.">Lung cancer and cryptogenic fibrosing alveolitis. A population-based cohort study.</a> Am J Resp Crit Care. 161, 5-8 (2000).</li><li>Nagai, A., Chiyotani, A., Nakadate, T., Konno, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lung+cancer+in+patients+with+idiopathic+pulmonary+fibrosis.">Lung cancer in patients with idiopathic pulmonary fibrosis.</a> Tohoku J Exp Med. 167, 231-237 (1992).</li><li>Rock, J. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+stromal+populations+contribute+to+pulmonary+fibrosis+without+evidence+for+epithelial+to+mesenchymal+transition.">Multiple stromal populations contribute to pulmonary fibrosis without evidence for epithelial to mesenchymal transition.</a> Proc Natl Acad Sci USA. 108, E1475-E1483 (2011).</li><li>Saito, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Survival+after+surgery+for+pathologic+stage+IA+non-small+cell+lung+cancer+associated+with+idiopathic+pulmonary+fibrosis.">Survival after surgery for pathologic stage IA non-small cell lung cancer associated with idiopathic pulmonary fibrosis.</a> Ann Thorac Surg. 92, 1812-1817 (2011).</li><li>Stevens, L. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extracellular+Matrix+Receptor+Expression+in+Subtypes+of+Lung+Adenocarcinoma+Potentiates+Outgrowth+of+Micrometastases.">Extracellular Matrix Receptor Expression in Subtypes of Lung Adenocarcinoma Potentiates Outgrowth of Micrometastases.</a> Cancer Res. 77, 1905-1917 (2017).</li><li>Harrison, J. H., Lazo, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+dose+continuous+infusion+of+bleomycin+in+mice:+a+new+model+for+drug-induced+pulmonary+fibrosis.">High dose continuous infusion of bleomycin in mice: a new model for drug-induced pulmonary fibrosis.</a> J Pharmacol Exp Ther. 243, 1185-1194 (1987).</li><li>Shcherbo, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bright+far-red+fluorescent+protein+for+whole-body+imaging.">Bright far-red fluorescent protein for whole-body imaging.</a> Nature Methods. 4, 741-746 (2007).</li><li>Aso, Y., Yoneda, K., Kikkawa, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Morphologic+and+biochemical+study+of+pulmonary+changes+induced+by+bleomycin+in+mice.">Morphologic and biochemical study of pulmonary changes induced by bleomycin in mice.</a> Lab Invest. 35, 558-568 (1976).</li><li>Kim, C. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+bronchioalveolar+stem+cells+in+normal+lung+and+lung.">Identification of bronchioalveolar stem cells in normal lung and lung.</a> Cell. 121, 823-835 (2005).</li><li>Fichtner, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishment+of+patient-derived+non-small+cell+lung+cancer+xenografts+as+models+for+the+identification+of+predictive+biomarkers.">Establishment of patient-derived non-small cell lung cancer xenografts as models for the identification of predictive biomarkers.</a> Clin Cancer Res. 14, 6456-6468 (2008).</li><li>Zhang, X. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishment+of+patient-derived+non-small+cell+lung+cancer+xenograft+models+with+genetic+aberrations+within+EGFR,+KRAS+and+FGFR1:+useful+tools+for+preclinical+studies+of+targeted+therapies.">Establishment of patient-derived non-small cell lung cancer xenograft models with genetic aberrations within EGFR, KRAS and FGFR1: useful tools for preclinical studies of targeted therapies.</a> J Transl Med. 11, 168 (2013).</li></ol>

Sorprendentes paralelismos clínicos han documentado entre cáncer de pulmón y otras enfermedades crónicas del pulmón36. En particular, los pacientes con fibrosis pulmonar idiopática (IPF) tienen una predilección mayor para desarrollar cáncer de pulmón, y esta asociación es independiente de fumar historia37,38. IPF se caracteriza por la destrucción progresiva de la arquitectura pulmonar y deterioro de la función respiratorio a tr...

Cáncer de pulmón es la principal causa del cáncer relacionadas con las muertes en todo el mundo1. Pacientes con cáncer de pulmón eventualmente sucumban de metástasis a órganos distantes, en particular para el sistema nervioso central, hígado, glándulas suprarrenales y huesos2,3,4. Tumores malignos torácicos han sido tradicionalmente clasificadas como cáncer de pulmón de células pequeñas (CPCP) o de células no pequeñas lung cancer (NSCLC)5. NSCLC es el más frecuentemente diagnosticado malignidad y puede subdividirse en diferentes subtipos histológicos, como el adenocarcinoma de pulmón (LUAD) y pulmón carcinoma de células escamosas (LUSC)6. Análisis genómico del cáncer de pulmón primario humano resecado ha revelado que los tumores dentro de un determinado histotype también pueden expresar diversas perturbaciones moleculares, contribuyendo a su evolución clínica divergente y confusión paciente pronóstico aún más. La notable heterogeneidad del cáncer de pulmón representa un desafío significativo al diseño racional, pruebas preclínicas y aplicación de estrategias terapéuticas eficaces. Por lo tanto, hay que ampliar el repertorio de modelos de cáncer de pulmón experimental manejable para estudiar los orígenes celulares diversos subtipos moleculares y etapas de esta enfermedad.
Diversos enfoques usando los modelos animales han sido empleados para el estudio de pulmón cáncer en vivo, cada uno con sus ventajas y desventajas dependiendo de las preguntas biológicas de interés. Modelos de ratón modificados genéticamente (GEMMs) pueden dirigirse a alteraciones genéticas específicas en un tipo de célula progenitora dado, resultante en tumores que progresan en un anfitrión inmunocompetente7. Aunque extremadamente potente y clínicamente relevantes, la morbosidad de tumor latencia, variabilidad o pulmón asociada GEMMs puede ser prohibitiva para algunas mediciones cuantitativas y la detección de metástasis de etapa tardía en órganos distantes8. Un enfoque complementario es el uso de modelos de aloinjerto, por el que las células cancerosas de pulmón, ya sea directamente de un tumor de ratón obtenidos o derivados primero como las líneas celulares establecidas en la cultura, son nuevamente introducidas en anfitriones syngeneic. Análogamente, xenoinjertos de cáncer de pulmón se establecen de líneas celulares humanas o muestras tumorales derivados pacientes. Xenoinjertos de línea celular humana o xenoinjertos paciente derivadas (PDXs) se mantienen generalmente en ratones inmunodeprimidos y por lo tanto excluye de vigilancia inmune completa9. A pesar de este inconveniente, que proporcionan una vía para propagar limitando cantidades de humanos hay y estudio fundamental en vivo propiedades de las células de cáncer en humanos, que codifican para aberraciones genómicas más complejas que los tumores GEMM.
Una propiedad útil de aloinjertos y xenoinjertos es que son susceptibles a la tradicional limitación celular dilución ensayos, empleados para cuantificar la frecuencia del tumor iniciar células (TICs) dentro de una población de células malignas10. En estos experimentos, un número definido de las células se inyecta por vía subcutánea en el flanco de los animales y la frecuencia de los TICs puede estimarse en base a la tasa de toma del tumor. Tumores subcutáneos sin embargo pueden ser más hipóxico11 y no pueden modelo claves limitaciones fisiológicas del microambiente del tumor de pulmón. Intratraqueal entrega de madre epitelial o células progenitoras en los pulmones de los ratones es un método para estudiar la regeneración pulmonar y las vías respiratorias la célula de vástago biología12. Sin embargo, la tasa de engraftment de esta técnica puede ser relativamente baja, a menos que los pulmones son sometidos primero a formas fisiológicas de lesiones, tales como infección viral13,14. Apoyo de las células stromal inflamatorias o la interrupción de la membrana basal de pulmón puede mejorar la retención de las células trasplantadas en nichos de células relevantes en la15de las vías aéreas distales. Agentes de inducción de fibrosis puede también previamente la condición los pulmones para mejorar el engraftment de células pluripotentes inducidas16 y17de las células madre mesenquimales. Si otras formas de lesión de las vías respiratorias pueden afectar la tasa de engraftment, capacidad de iniciación tumoral y consecuencia de las células de cáncer de pulmón todavía debe evaluarse sistemáticamente.
En este estudio, se describe un método para aumentar la eficiencia de orthotopic engraftment de la célula de cáncer de pulmón, por acondicionamiento previo de los pulmones de los ratones con lesión. LUAD se presenta en las vías aéreas distales con un subconjunto significativo de estos cánceres en el desarrollo de un tejido conectador fibrótico18 que a menudo se correlaciona con mal pronóstico19. Bleomicina, un peptide nonribosomal natural del híbrido-polyketide, ha sido ampliamente utilizada para inducir fibrosis pulmonar en ratones20. Instilación de las vías respiratorias de la bleomicina primero promueve el desgaste epitelial de los alvéolos y el reclutamiento de células inflamatorias, incluyendo macrófagos, neutrófilos y monocitos21. Esto es seguido por remodelación tisular en las vías aéreas distales, reorganización de la membrana del sótano22,23 y deposición de matriz extracelular (ECM)24. Los efectos de una inyección de bleomicina solo son transitorios, con fibrosis resolver después de 30 días en la mayoría de los estudios de25. Utilizando modelos de aloinjertos y xenoinjertos, probamos si acondicionamiento previo de las vías respiratorias de los ratones con bleomicina puede aumentar significativamente el índice de toma de LUAD células en los pulmones.

<table>
	<tbody>
		<tr>
			<td>Bleomycin</td>
			<td>Sigma</td>
			<td>B5507-15UN</td>
			<td>CAUTION Health hazard GHS08</td>
		</tr>
		<tr>
			<td>Exel Catheter 24G</td>
			<td>Fisher</td>
			<td>1484121</td>
			<td>Remove needle. For intratracheal injection</td>
		</tr>
		<tr>
			<td>Ketamine (Ketaset inl 100 mg/mL C3N 10 mL)</td>
			<td>Butler Schein</td>
			<td>56344</td>
			<td>To anesthetize mice</td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>Butler Schein</td>
			<td>33198</td>
			<td>To anesthetize mice</td>
		</tr>
		<tr>
			<td>Ketoprofen, 5,000 mg</td>
			<td>Cayman Chemical</td>
			<td>10006661</td>
			<td>Analgesic</td>
		</tr>
		<tr>
			<td>Puralube Veterinary Ophthalmic Ointment</td>
			<td>BUTLER ANIMAL HEALTH COMPANY LLC</td>
			<td>8897</td>
			<td>To prevent eye dryness while under anesthesia</td>
		</tr>
		<tr>
			<td>D-Luciferin powder</td>
			<td>Perkin Elmer Health Sciences Inc</td>
			<td>122799</td>
			<td>For luminescent imaging. Reconstitute powder with PBS for a working concentration of 15mg/mL. Protect from Light</td>
		</tr>
		<tr>
			<td>Rodent Intubation stand</td>
			<td>Braintree Scientific</td>
			<td>RIS-100</td>
			<td>Recommended stand for intratracheal injection</td>
		</tr>
		<tr>
			<td>MI-150 ILLUMINATOR 150W MI-150</td>
			<td>DOLAN-JENNER INDUSTRIES</td>
			<td>MI-150 / EEG2823M</td>
			<td>To illuminate and visualize trachea</td>
		</tr>
		<tr>
			<td>Graefe Forceps, 2.75 (7 cm) long serrat</td>
			<td>Roboz</td>
			<td>RS-5111</td>
			<td>For intratracheal injection</td>
		</tr>
		<tr>
			<td>Syringe Luer-Lok Sterile 5ml</td>
			<td>BD / Fisher</td>
			<td>309646</td>
			<td></td>
		</tr>
		<tr>
			<td>Satiny Smooth by Conair Dual Foil Wet/Dry Rechargeable Shaver</td>
			<td>Conair</td>
			<td>-</td>
			<td>To shave mice</td>
		</tr>
		<tr>
			<td>Bonn Scissors, 3.5&#34; straight 15 mm sharp/sharp sure cut blades</td>
			<td>Roboz</td>
			<td>RS-5840SC</td>
			<td></td>
		</tr>
		<tr>
			<td>15 mL conical tube</td>
			<td>BD / Fisher</td>
			<td>352097</td>
			<td></td>
		</tr>
		<tr>
			<td>1.5 mL centrifuge tubes</td>
			<td>USA SCIENTIFIC INC</td>
			<td>1615-5500</td>
			<td></td>
		</tr>
		<tr>
			<td>Vial Scintillation 7 mL Borosilicate Glass GPI</td>
			<td>Fisher</td>
			<td>701350</td>
			<td></td>
		</tr>
		<tr>
			<td>Filter pipette tips (200 &#956;L)</td>
			<td>USA SCIENTIFIC INC</td>
			<td>1120-8710</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate Buffered Saline</td>
			<td>Life Technologies</td>
			<td>14190-144</td>
			<td></td>
		</tr>
		<tr>
			<td>0.25% Trypsin-EDTA</td>
			<td>Life Technologies</td>
			<td>25200-056</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM high glucose</td>
			<td>Life Technologies</td>
			<td>11965-092</td>
			<td></td>
		</tr>
		<tr>
			<td>RPMI Medium 1640</td>
			<td>Life Technologies</td>
			<td>11875-093</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal bovine serum USDA</td>
			<td>Life Technologies</td>
			<td>10437-028</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin-Streptomycin</td>
			<td>Life Technologies</td>
			<td>15140-122</td>
			<td></td>
		</tr>
		<tr>
			<td>Amphotericin B</td>
			<td>Sigma</td>
			<td>A2942-20ML</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypan Blue Stain 0.4%</td>
			<td>Life Technologies</td>
			<td>15250-061</td>
			<td></td>
		</tr>
		<tr>
			<td>Countess Automated Cell Counter</td>
			<td>Life Technologies</td>
			<td>AMQAX1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Flask T/C 75cm sq canted neck, blue cap</td>
			<td>Fisher / Corning</td>
			<td>353135</td>
			<td></td>
		</tr>
		<tr>
			<td>IVIS Spectrum Xenogen Bioluminiscence</td>
			<td>Perkin Elmer Health Sciences Inc</td>
			<td>124262</td>
			<td>For in vivo bioluminescence imaging</td>
		</tr>
		<tr>
			<td>Living image software</td>
			<td>Perkin Elmer Health Sciences Inc</td>
			<td>128113</td>
			<td>For in vivo bioluminescence analysis</td>
		</tr>
		<tr>
			<td>XGI-8 Gas Anesthesia System</td>
			<td>Perkin Elmer Health Sciences Inc</td>
			<td>118918</td>
			<td>For Isoflurane anesthesia</td>
		</tr>
		<tr>
			<td>BD Ultra-Fine II Short Needle Insulin Syringe 1 cc. 31 G x 8 mm (5/16 in)</td>
			<td>BD / Fisher</td>
			<td>BD328418</td>
			<td>For retro-orbital luciferin injection</td>
		</tr>
		<tr>
			<td>Syringe 1ml</td>
			<td>BD / Fisher</td>
			<td>14-823-434</td>
			<td>For intraperitoneal injections</td>
		</tr>
		<tr>
			<td>26 G x 1/2 in. needle</td>
			<td>BD / Fisher</td>
			<td>305111</td>
			<td>For intraperitoneal injections</td>
		</tr>
		<tr>
			<td>4% Paraformaldehyde</td>
			<td>VWR</td>
			<td>43368-9M</td>
			<td>CAUTION Health hazard GHS07, GHS08. For fixing tissue</td>
		</tr>
		<tr>
			<td>Pipet-Lite Pipette, Unv. SL-200XLS+</td>
			<td>METTLER-TOLEDO INTERNATIONAL</td>
			<td>17014411</td>
			<td></td>
		</tr>
		<tr>
			<td>Mayer&#39;s Hematoxylin</td>
			<td>ELECTRON MICROSCOPY SCIENCES</td>
			<td>517-28-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Eosin Y stain 0.25% (w/v) in 57%</td>
			<td>Fisher</td>
			<td>67-63-0</td>
			<td></td>
		</tr>
		<tr>
			<td>Masson Trichrome Stain Kit</td>
			<td>IMEB Inc</td>
			<td>K7228</td>
			<td>For masson trichrome stain to visualize collagen</td>
		</tr>
		<tr>
			<td>Superfrost plus glass slides</td>
			<td>Fisher</td>
			<td>1255015</td>
			<td></td>
		</tr>
		<tr>
			<td>6 well plate</td>
			<td>Corning</td>
			<td>C3516</td>
			<td></td>
		</tr>
		<tr>
			<td>Universal Mycoplasma Detection Kit</td>
			<td>ATCC</td>
			<td>30-1012K</td>
			<td></td>
		</tr>
		<tr>
			<td>OCT Embedding compound</td>
			<td>ELECTRON MICROSCOPY SCIENCES</td>
			<td>62550-12</td>
			<td>For embedding tissue for frozen sections</td>
		</tr>
		<tr>
			<td>Leica CM3050 S Research Cryostat</td>
			<td>Leica</td>
			<td>CM3050 S</td>
			<td>To section tissue for staining analysis</td>
		</tr>
		<tr>
			<td>Keyence All-in One Fluorescence Microscope</td>
			<td>Keyence</td>
			<td>BZ-X700</td>
			<td></td>
		</tr>
		<tr>
			<td>ImageJ</td>
			<td>US National Institutes of Health</td>
			<td>IJ1.46</td>
			<td>http://rsbweb.nih.gov/ij/ download.html</td>
		</tr>
		<tr>
			<td>Prism 7.0 for Mac OS X</td>
			<td>GraphPad Software, Inc.</td>
			<td>-</td>
			<td></td>
		</tr>
		<tr>
			<td>Athymic (Crl:NU(NCr)-Foxn1nu) mice</td>
			<td>Charles River</td>
			<td>NIH-553</td>
			<td></td>
		</tr>
		<tr>
			<td>NSG (NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ) mice</td>
			<td>Jackson Laboratories</td>
			<td>5557</td>
			<td></td>
		</tr>
		<tr>
			<td>B6129SF1/J mice</td>
			<td>Jackson Laboratories</td>
			<td>101043</td>
			<td></td>
		</tr>
		<tr>
			<td>NIH-H2030 cells</td>
			<td>ATCC</td>
			<td>CRL-5914</td>
			<td></td>
		</tr>
		<tr>
			<td>368T1</td>
			<td>generously provided by Monte Winslow (Standford University)</td>
			<td>-</td>
			<td></td>
		</tr>
		<tr>
			<td>PC9 cells</td>
			<td>Nguyen DX et al. Cell. 2009;138:51&#8211;62</td>
			<td>-</td>
			<td></td>
		</tr>
		<tr>
			<td>H2030 BrM3 cells</td>
			<td>Nguyen DX et al. Cell. 2009;138:51&#8211;62</td>
			<td>-</td>
			<td></td>
		</tr>
	</tbody>
</table>

pre-conditioning the airways of mice with bleomycin increases the efficiency of orthotopic lung cancer cell engraftment

Cáncer de pulmón es una enfermedad refractaria mortal tratamiento biológico heterogéneo. Para entender y tratar con eficacia el espectro clínico completo de tumores malignos torácicos, se necesitan modelos animales adicionales que pueden recapitular fases y subtipos de cáncer de pulmón humanas diversas. Aloinjerto o xenoinjerto modelos son versátiles y permiten la cuantificación de capacidad tumorígena en vivo, usando las células malignas de origen murino o humano. Sin embargo, se han realizado métodos previamente descritos del engraftment de la célula de pulmón cáncer en sitios no-fisiológicos, tales como el flanco de los ratones, debido a la ineficiencia del trasplante orthotopic de células en los pulmones. En este estudio, se describe un método para mejorar el acondicionamiento previo de las vías respiratorias de los ratones con la bleomicina de agente induce fibrosis orthotopic engraftment de la célula de cáncer de pulmón. Como un experimento de prueba de concepto, se aplicó este enfoque para engraft las células del tumor del subtipo de adenocarcinoma de pulmón, obtenida de fuentes humanas, o de ratón en varias cepas de ratones. Demostramos que hiriendo a las vías respiratorias con bleomicina antes de la inyección de células de tumor aumenta el engraftment de células tumorales de 0-17% 71-100%. Este método había mejorado significativamente, incidencia de tumor de pulmón y posterior consecuencia utilizando cepas de ratón y diferentes modelos. Además, las células de cáncer de pulmón engrafted difusión desde los pulmones a órganos distantes pertinentes. Por lo tanto, proporcionamos un protocolo que puede utilizarse para establecer y mantener nuevos modelos ortotópico de cáncer de pulmón con limitación de la cantidad de células o muestra biológica y evaluar cuantitativamente la capacidad tumorigénica de las células de cáncer de pulmón en configuración fisiológico correspondiente .

Para aumentar la eficiencia del engraftment de la célula cáncer LUAD en los pulmones de ratones, hemos desarrollado un protocolo que primero las condiciones de las vías respiratorias utilizando bleomicina seguida de la inyección de células de tumor ortotópico (figura 1). Se confirmó que incluso cuando se administra en ratones athymic immunocompromised, bleomicina induce fibrosis transitoria día 14 según lo evidenciado por la pérdida de la arquitectu...

Watch this Scientific Journal Video about Pre-Conditioning the Airways of Mice with Bleomycin Increases the Efficiency of Orthotopic Lung Cancer Cell Engraftment at JoVE.com

Pre-Conditioning the Airways of Mice with Bleomycin Increases the Efficiency of Orthotopic Lung Cancer Cell Engraftment

Se describe un método para mejorar considerablemente orthotopic engraftment de células de cáncer de pulmón en los pulmones murinos por acondicionamiento previo de las vías respiratorias con lesiones. Este enfoque también puede aplicarse para estudiar las interacciones estromales dentro del microambiente de la pulmonar, diseminación metastásica, comorbilidades de cáncer de pulmón, y generar más eficientemente paciente derivados de xenoinjertos.

Acondicionamiento previo de las vías respiratorias de los ratones con bleomicina aumenta la eficacia del pulmón Orthotopic Engraftment de la célula de cáncer

Lung cancer is a deadly treatment refractory disease that is biologically heterogeneous. To understand and effectively treat the full clinical spectrum of ...

The overall goal of this procedure is to increase the efficiency of transplanting tumor-initiating lung cancer cells into the airways of mice. This method can help establish new preclinical models in order to answer key questions in cancer biology, including the causes of drug resistance and relationship between fibrosis and lung cancer. The main advantage of this technique is that it increase the transplantation efficiency of tumor-initiating cells into the lungs of a variety of mouse strains.The implications of this technique extend toward therapy, because it enables us to model a greater variety of human lung cancer subtypes, known to have different clinical responses to treatment. For this protocol, the mice must first be injected with bleomycin. Then, after 14 days, the tumor cells are injected.The procedure for both injections is very similar and described here. Two hours prior to the injection, thaw the bleomycin stock on ice and dilute it to the working concentration. Keep it on ice.For a cell bolus, have a cell suspension prepared as described in the text protocol. The procedure time for each mouse is about 10 minutes all together. Be certain to confirm proper anesthesia using a toe pinch, and apply vet ointment to the eyes.At this time, if the mouse has dark hair, then shave the entire rib area both ventrally and dorsally so it can be imaged correctly. Then, place the mouse on the intubation platform by securing it by its front teeth with it's back against the platform. If a cell bolus is to be injected, resuspend it now with gentle trituration.Now illuminate the upper-chest region, open the mouth, and gently pull out the tongue using flat forceps. Prepare an IV catheter for the trachea by removing the needle, then position it over the tracheal opening. Next, insert the catheter into the trachea until the top of the catheter is near the front teeth and illuminate the interior of the catheter to confirm its placement.Then, dispense the 50 microliters of freshly made bleomycin, or cell suspension, directly into the catheter. Wait a few seconds for the entire volume to travel down the catheter. Then, remove the catheter and dispose of it in 10%bleach.If the mouse is not inhaling the liquid, carefully monitor its breathing and adjust the catheter position. If the mouse stops breathing, immediately remove the catheter and allow the mouse to breath before you reinsert the catheter in. After removing the catheter, continue performing the procedure to have five treated mice, still anesthetized, and ready to image.For recovery, place the mouse supine on an IACUC approved heating pad in a recovery cage and monitor it until it has fully recovered. Two or three minutes after injecting the cells, inject 100 microliters of luciferin retro-orbitally, using an insulin needle. Alternate the eye used for injection every imaging session.After waiting at lest two minutes, position up to five mice in an animal bioluminescence imager, in the sternal recumbency position and acquire a dorsal picture using luminescence settings. Under the control panel, adjust the resolution and sensitivity settings, to measure the luminescence of the cells. For most applications, start with three minutes, set the binning to medium, set the f/stop to one, set the field of view to d, and set the subject height to 1.5.If the injection was successful, a signal is detected in the upper chest area in one lung or both lungs. If cells are clustered at the entrance of the trachea, the injection was unsuccessful. After taking the dorsal image, flip the mice and acquire a ventral image and adjust the setting as needed.After imaging, proceed with postoperative care as described in the text protocol. Then, repeat this imaging protocol three days post-engraftment and weekly thereafter. To analyze the images, use a bioluminescence analysis software.First, load all the appropriate images in the series. This section is detailed to one software package. Next, for each mouse, in the first image of the series, create a square region of interest and position it over the upper chest area covering the entire lung region.Then, using a right click, access the copy all regions of interest function, and use this tool to apply the same regions of interest to each image in the group. Then, within each image, tweak the positions of the squares to cover the upper chest of each mouse. Do this for both the ventral and dorsal views.Now, in the upper left corner of the image window, select the photons function. Then select the measure option. The data from the regions of interest is then displayed in an output table as total flux.Analyze this data as needed. When immune compromised, athymic mice were treated with bleomycin. Prior to cell-bolus engraftment, transient fibrosis was observed by day 14 in bleomycin-treated mice, but not vehicle-treated mice.Next, cells from the established human lung cancer cell line, H2030, were delivered intratracheally into the lungs of these mice. After 35 days, bleomycin-treated mice had a high lung-tumor burden. However, in vehicle treated animals, no tumors were detected.The lung-tumor burden was confirmed using histology. Lungs pretreated with vehicle had no evidence of tumor nodules, whereas bleomycin pretreated lungs had large tumor nodules. After transplanting a metastatic H2030 BrM 3 cell line, by 39 to 57 days, bioluminescence was too intense for imaging, so metastasis was analyzed in distant organs ex vivo.Small lesions could be detected in various tissues, suggesting that this method holds promise, to study spontaneously disseminated disease. Once mastered, this technique can be done in 10 minutes per cage of five mice without imaging and 20 minutes per cage when also imaging the mice. While attempting this procedure, it is important to remember to ensure proper insertion of the catheter and to monitor the animal's breathing.Following this procedure, other methods like pharmacological studies or aminoacetic chemistry of harvested tissue can be performed in order to answer additional questions, like how engrafted tumors respond to drugs or characterizing tumor stromal interactions specific to the lungs. After its development, this technique can be applied to different mouse strains and tumor models, including patient-derived xenografts in a physiologically relevant setting. Don't forgot that working with bleomycin can be extremely hazardous and precautions such as manipulation in a biohazard hood and proper disposal should always be taken while performing this procedure.

Research

JoVE Journal

Medicine

Bronchial Thermoplasty:  A Novel Therapeutic Approach to Severe Asthma

Bronquial termoplastia: un nuevo enfoque terapéutico para el asma grave

Bronchial thermoplasty is a non-drug procedure for severe persistent asthma that delivers thermal energy to the airway wall in a precisely controlled ...

Investigación

Medicina

An Orthotopic Model of Murine Bladder Cancer

Un modelo ortotópico de cáncer de vejiga murino

In this straightforward procedure, bladder tumors are established in female C57 mice through the use of catheterization, local cauterization, and ...

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

Multi-fotón imágenes de invasión de células tumorales en un modelo de ratón ortotópico de carcinoma oral de células escamosas

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

An Orthotopic Bladder Tumor Model and the Evaluation of Intravesical saRNA Treatment

Un modelo de tumor vesical ortotópica y la Evaluación de tratamiento intravesical SARNA

We present a novel method for treating bladder cancer with intravesically delivered small activating RNA (saRNA) in an orthotopic xenograft mouse bladder ...

Orthotopic Aortic Transplantation in Mice for the Study of Vascular Disease

El trasplante ortotópico aórtica en los ratones para el Estudio de la Enfermedad Vascular

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. ...

A Murine Model of Myocardial Ischemia-reperfusion Injury through Ligation of the Left Anterior Descending Artery

Un modelo murino de isquemia-reperfusión miocárdica lesión a través de la ligadura de la arteria descendente anterior

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms ...

Orthotopic Injection of Breast Cancer Cells into the Mammary Fat Pad of Mice to Study Tumor Growth.

Inyección ortotópico de células de cáncer de mama en el Mamaria Fat Pad de ratones para estudiar el crecimiento del tumor.

Breast cancer growth can be studied in mice using a plethora of models. Genetic manipulation of breast cancer cells may provide insights into the ...

Non-invasive Assessment of the Efficacy of New Therapeutics for Intestinal Pathologies Using Serial Endoscopic Imaging of Live Mice

Evaluación no invasiva de la eficacia de nuevas terapias para Intestinal Patologías recurriendo a imágenes de Endoscópica serie de ratones vivos

Animal models of inflammatory bowel disease (IBD) and colorectal cancer (CRC) have provided significant insight into the cell intrinsic and extrinsic ...

Cardiac Catheterization in Mice to Measure the Pressure Volume Relationship: Investigating the Bowditch Effect

Cateterismo cardíaco en ratones para medir la relación de volumen Presión: Investigar el efecto Bowditch

Animal models that mimic human cardiac disorders have been created to test potential therapeutic strategies. A key component to evaluating these ...

A "Patient-Like" Orthotopic Syngeneic Mouse Model of Hepatocellular Carcinoma Metastasis

Un &quot;paciente-Like&quot; ortotópico Singénico modelo de ratón de carcinoma hepatocelular Metástasis

The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of ...