{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T15:22:34Z","timestamp":1787239354800,"version":"build-2736575974"},"reference-count":42,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2020,12,2]],"date-time":"2020-12-02T00:00:00Z","timestamp":1606867200000},"content-version":"vor","delay-in-days":1,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["currentprotocols.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["CP Pharmacology"],"published-print":{"date-parts":[[2020,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Cancer\u2010associated cachexia is defined by loss of weight and muscle mass, and by the potential loss of adipose tissue accompanied by insulin resistance and increased resting energy expenditure. Cachexia is most prevalent in pancreatic cancer, the third leading cause of cancer\u2010related deaths. While various factors interact to induce cachexia, the precise mechanisms underlying this clinical condition are not fully understood. Clinically relevant animal models of cachexia are needed given the lack of standard diagnostic methods or treatments for this condition. Described in this article are in vitro and in vivo models used to study the role of macrophages in the induction of cachexia in pancreatic cancer. Included are procedures for isolating and culturing bone marrow\u2212derived macrophages, harvesting tumor\u2010 and macrophage\u2010derived conditioned medium, and studying the effect of conditioned medium on C2C12 myotubes. Also described are procedures involving the use of an orthotopic model of pancreatic cancer, including a method for examining skeletal muscle atrophy in this model. \u00a9 2020 Wiley Periodicals LLC.<\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 1<\/jats:bold>\n                    : In vitro model of pancreatic tumor\u2010induced cachexia using C2C12 cell lines (myotube model)\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 1<\/jats:bold>\n                    : Molecular evaluation of cachectic markers in C2C12 myotubes using real\u2010time PCR and immunoblotting\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 2<\/jats:bold>\n                    : In vivo model to study cachectic phenotype in pancreatic tumor\u2010bearing mice\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 2<\/jats:bold>\n                    : Evaluation of cachectic markers in the skeletal muscle of tumor\u2010bearing mice\n                  <\/jats:p>","DOI":"10.1002\/cpph.80","type":"journal-article","created":{"date-parts":[[2020,12,2]],"date-time":"2020-12-02T17:27:35Z","timestamp":1606930055000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Preclinical Models for Studying the Impact of Macrophages on Cancer Cachexia"],"prefix":"10.1002","volume":"91","author":[{"given":"Spas Dimitrov","family":"Markov","sequence":"first","affiliation":[{"name":"The Eppley Institute for Cancer and Allied Diseases University of Nebraska Medical Center  Omaha Nebraska"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daisy","family":"Gonzalez","sequence":"additional","affiliation":[{"name":"The Eppley Institute for Cancer and Allied Diseases University of Nebraska Medical Center  Omaha Nebraska"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kamiya","family":"Mehla","sequence":"additional","affiliation":[{"name":"The Eppley Institute for Cancer and Allied Diseases University of Nebraska Medical Center  Omaha Nebraska"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2020,12,2]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12885-018-5080-4"},{"key":"e_1_2_7_3_1","doi-asserted-by":"crossref","first-page":"17\u221253","DOI":"10.1002\/9780470942390.mo100118","article-title":"Standardized post\u2010mortem examination and fixation procedures for mutant and treated mice","volume":"1","author":"Antal C.","year":"2011","journal-title":"Current Protocols in Mouse Biology"},{"key":"e_1_2_7_4_1","doi-asserted-by":"crossref","first-page":"2408","DOI":"10.1038\/s41598-018-20820-4","article-title":"Microscale Gene Expression Analysis of Tumor\u2010Associated Macrophages","volume":"8","author":"Attri K. S.","year":"2018","journal-title":"Scientific Reports"},{"key":"e_1_2_7_5_1","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1007\/978-1-4939-7665-2_23","article-title":"Evaluation of macrophage polarization in pancreatic cancer microenvironment under hypoxia","volume":"1742","author":"Attri K. S.","year":"2018","journal-title":"Methods in Molecular Biology"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1065874"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.95.25.15043"},{"key":"e_1_2_7_8_1","first-page":"223","article-title":"C2C12 murine myoblasts as a model of skeletal muscle development: Morpho\u2010functional characterization","volume":"48","author":"Burattini S.","year":"2004","journal-title":"European Journal of Histochemistry"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/nn1455"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1302895"},{"key":"e_1_2_7_11_1","first-page":"A.2A.1\u2212A.2A.3","article-title":"Common stock solutions, buffer, and media","volume":"00","author":"Current Protocols","year":"2001","journal-title":"Current Protocols in Pharmacology"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20190745"},{"key":"e_1_2_7_13_1","first-page":"1.4.1\u22121.4.5","article-title":"Anesthesia","volume":"27","author":"Donovan J.","year":"2001","journal-title":"Current Protocols in Immunology"},{"key":"e_1_2_7_14_1","doi-asserted-by":"crossref","first-page":"1.6.1\u22121.6.10","DOI":"10.1002\/0471142735.im0106s73","article-title":"Parenteral injections","volume":"73","author":"Donovan J.","year":"2006","journal-title":"Current Protocols in Immunology"},{"key":"e_1_2_7_15_1","doi-asserted-by":"crossref","first-page":"1.8.1\u22121.8.4","DOI":"10.1002\/0471142735.im0108s73","article-title":"Euthanasia","volume":"73","author":"Donovan J.","year":"2006","journal-title":"Current Protocols in Immunology"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.2353\/ajpath.2010.090884"},{"key":"e_1_2_7_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1470-2045(10)70218-7"},{"key":"e_1_2_7_18_1","doi-asserted-by":"publisher","DOI":"10.1002\/0471143030.cb0601s37"},{"key":"e_1_2_7_19_1","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-16-3099"},{"key":"e_1_2_7_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2007.10.003"},{"key":"e_1_2_7_21_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.251541198"},{"key":"e_1_2_7_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.physbeh.2010.03.011"},{"key":"e_1_2_7_23_1","doi-asserted-by":"publisher","DOI":"10.1097\/MPA.0000000000001124"},{"key":"e_1_2_7_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccr.2005.04.023"},{"key":"e_1_2_7_25_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrc3972"},{"key":"e_1_2_7_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.gtc.2007.08.012"},{"key":"e_1_2_7_27_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep44570"},{"key":"e_1_2_7_28_1","doi-asserted-by":"publisher","DOI":"10.1006\/meth.2001.1262"},{"key":"e_1_2_7_29_1","doi-asserted-by":"publisher","DOI":"10.3389\/fonc.2020.00298"},{"key":"e_1_2_7_30_1","first-page":"1432","article-title":"Role of the central melanocortin system in cachexia","volume":"61","author":"Marks D. L.","year":"2001","journal-title":"Cancer Research"},{"key":"e_1_2_7_31_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/978-1-62703-287-2_1","article-title":"Identification and analysis of precursors to invasive pancreatic cancer","volume":"980","author":"Matthaei H.","year":"2013","journal-title":"Methods in Molecular Biology"},{"key":"e_1_2_7_32_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpcell.1994.266.6.C1795"},{"key":"e_1_2_7_33_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcsm.12225"},{"key":"e_1_2_7_34_1","doi-asserted-by":"publisher","DOI":"10.1242\/dmm.008839"},{"key":"e_1_2_7_35_1","first-page":"6.2.1\u22126.2.37","article-title":"Immunoblotting and immunodetection","volume":"74","author":"Ni D.","year":"2017","journal-title":"Current Protocols in Cell Biology"},{"key":"e_1_2_7_36_1","doi-asserted-by":"crossref","first-page":"12.1.1\u221212.1.23","DOI":"10.1002\/cpph.1","article-title":"Mammalian cell tissue culture techniques","volume":"73","author":"Phelan K.","year":"2016","journal-title":"Current Protocols in Pharmacology"},{"key":"e_1_2_7_37_1","doi-asserted-by":"publisher","DOI":"10.18632\/oncotarget.5843"},{"key":"e_1_2_7_38_1","doi-asserted-by":"publisher","DOI":"10.1186\/2049-3002-2-18"},{"key":"e_1_2_7_39_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.canlet.2020.04.017"},{"key":"e_1_2_7_40_1","article-title":"Macrophages potentiate STAT3 signaling in skeletal muscles and regulate pancreatic cancer cachexia","author":"Shukla S. K.","year":"2020","journal-title":"Cancer Letters"},{"key":"e_1_2_7_41_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2019.07.016"},{"key":"e_1_2_7_42_1","doi-asserted-by":"publisher","DOI":"10.1038\/270725a0"},{"key":"e_1_2_7_43_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcsm.12461"}],"container-title":["Current Protocols in Pharmacology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpph.80","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/cpph.80","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpph.80","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,3]],"date-time":"2023-09-03T05:01:39Z","timestamp":1693717299000},"score":1,"resource":{"primary":{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/10.1002\/cpph.80"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,12]]},"references-count":42,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2020,12]]}},"alternative-id":["10.1002\/cpph.80"],"URL":"https:\/\/doi.org\/10.1002\/cpph.80","archive":["Portico"],"relation":{},"ISSN":["1934-8282","1934-8290"],"issn-type":[{"value":"1934-8282","type":"print"},{"value":"1934-8290","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,12]]},"assertion":[{"value":"2020-12-02","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e80"}}