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{"title":"胰腺导管类器官的生成及完整类器官的全载免疫染色","authors":"Habib Rezanejad, Jennifer Hollister Lock, Brooke A Sullivan, Susan Bonner-Weir","doi":"10.1002/cpcb.82","DOIUrl":null,"url":null,"abstract":"<p>Traditionally, studies of cells and tissues have been performed on isolated primary cells or immortalized cell lines by culturing them in 2D culture dishes or flasks. However, a caveat regarding 2D culture is that the cells poorly recapitulate their <i>in vivo</i> counterparts, mainly due to a lack of 3D cell-cell and cell–extracellular matrix interactions. In recent years, the development of <i>in vitro</i> organoids as 3D culture has gained substantial attention as a model to study different tissues. In adults, pancreatic ductal cells are considered as a source of stem or progenitor cells, so developing new methods to study ductal cells would be useful. Here, we provide a protocol to isolate mouse pancreatic ductal cells and a cost-effective protocol to generate 3D organoid structures from such ductal cells. Additionally, we have devised a protocol for immunostaining of intact whole organoids without sectioning. © 2018 by John Wiley & Sons, Inc.</p>","PeriodicalId":40051,"journal":{"name":"Current Protocols in Cell Biology","volume":"83 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpcb.82","citationCount":"7","resultStr":"{\"title\":\"Generation of Pancreatic Ductal Organoids and Whole-Mount Immunostaining of Intact Organoids\",\"authors\":\"Habib Rezanejad, Jennifer Hollister Lock, Brooke A Sullivan, Susan Bonner-Weir\",\"doi\":\"10.1002/cpcb.82\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Traditionally, studies of cells and tissues have been performed on isolated primary cells or immortalized cell lines by culturing them in 2D culture dishes or flasks. However, a caveat regarding 2D culture is that the cells poorly recapitulate their <i>in vivo</i> counterparts, mainly due to a lack of 3D cell-cell and cell–extracellular matrix interactions. In recent years, the development of <i>in vitro</i> organoids as 3D culture has gained substantial attention as a model to study different tissues. In adults, pancreatic ductal cells are considered as a source of stem or progenitor cells, so developing new methods to study ductal cells would be useful. Here, we provide a protocol to isolate mouse pancreatic ductal cells and a cost-effective protocol to generate 3D organoid structures from such ductal cells. Additionally, we have devised a protocol for immunostaining of intact whole organoids without sectioning. © 2018 by John Wiley & Sons, Inc.</p>\",\"PeriodicalId\":40051,\"journal\":{\"name\":\"Current Protocols in Cell Biology\",\"volume\":\"83 1\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2018-12-12\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1002/cpcb.82\",\"citationCount\":\"7\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Current Protocols in Cell Biology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/cpcb.82\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"Biochemistry, Genetics and Molecular Biology\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Cell Biology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpcb.82","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
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