Bianca Marigliani, Luciene Bottentuit López Balottin, Elisabeth de Fatima Pires Augusto
{"title":"哺乳动物细胞对化学介质的适应","authors":"Bianca Marigliani, Luciene Bottentuit López Balottin, Elisabeth de Fatima Pires Augusto","doi":"10.1002/cptx.88","DOIUrl":null,"url":null,"abstract":"<p>In order to circumvent ethical, technical, and economic drawbacks regarding the use of animal serum in cell culturing, it is possible to adapt mammalian cells to serum-free media. Nowadays, there are several serum-free formulations available, including fully animal derived–free and chemically defined media, and different adaptation techniques. This article focuses on the gradual adaptation of a mammalian suspension cell culture to a chemically defined medium. The first step is to transfer the cells cultured in medium supplemented with fetal bovine serum (FBS) to a chemically defined medium of your choice, containing the same amount of FBS. The next steps consist of progressively reducing the amount of FBS, while monitoring cell growth and viability up to the complete elimination of FBS. This protocol has been successfully used to adapt THP-1 cells to a chemically defined medium with similar maximum specific growth rate as those cultured with FBS. © 2019 by John Wiley & Sons, Inc.</p><p><b>Basic Protocol</b>: Gradual adaptation to chemically defined medium</p><p><b>Alternate Protocol</b>: Direct adaptation to chemically defined medium</p><p><b>Support Protocol 1</b>: Determining maximum specific growth rate of a cell culture</p><p><b>Support Protocol 2</b>: Cell freezing and thawing</p>","PeriodicalId":72743,"journal":{"name":"Current protocols in toxicology","volume":"82 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2019-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cptx.88","citationCount":"10","resultStr":"{\"title\":\"Adaptation of Mammalian Cells to Chemically Defined Media\",\"authors\":\"Bianca Marigliani, Luciene Bottentuit López Balottin, Elisabeth de Fatima Pires Augusto\",\"doi\":\"10.1002/cptx.88\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>In order to circumvent ethical, technical, and economic drawbacks regarding the use of animal serum in cell culturing, it is possible to adapt mammalian cells to serum-free media. Nowadays, there are several serum-free formulations available, including fully animal derived–free and chemically defined media, and different adaptation techniques. This article focuses on the gradual adaptation of a mammalian suspension cell culture to a chemically defined medium. The first step is to transfer the cells cultured in medium supplemented with fetal bovine serum (FBS) to a chemically defined medium of your choice, containing the same amount of FBS. The next steps consist of progressively reducing the amount of FBS, while monitoring cell growth and viability up to the complete elimination of FBS. This protocol has been successfully used to adapt THP-1 cells to a chemically defined medium with similar maximum specific growth rate as those cultured with FBS. © 2019 by John Wiley & Sons, Inc.</p><p><b>Basic Protocol</b>: Gradual adaptation to chemically defined medium</p><p><b>Alternate Protocol</b>: Direct adaptation to chemically defined medium</p><p><b>Support Protocol 1</b>: Determining maximum specific growth rate of a cell culture</p><p><b>Support Protocol 2</b>: Cell freezing and thawing</p>\",\"PeriodicalId\":72743,\"journal\":{\"name\":\"Current protocols in toxicology\",\"volume\":\"82 1\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2019-10-10\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1002/cptx.88\",\"citationCount\":\"10\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Current protocols in toxicology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/cptx.88\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current protocols in toxicology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cptx.88","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 10