Bianca Marigliani, Luciene Bottentuit López Balottin, Elisabeth de Fatima Pires Augusto
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引用次数: 10
Abstract
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.
Basic Protocol: Gradual adaptation to chemically defined medium
Alternate Protocol: Direct adaptation to chemically defined medium
Support Protocol 1: Determining maximum specific growth rate of a cell culture
Support Protocol 2: Cell freezing and thawing
哺乳动物细胞对化学介质的适应
为了避免在细胞培养中使用动物血清的伦理、技术和经济缺陷,有可能使哺乳动物细胞适应无血清培养基。如今,有几种无血清制剂可供选择,包括完全不含动物源性和化学定义的培养基,以及不同的适应技术。这篇文章的重点是逐渐适应哺乳动物悬浮细胞培养到一种化学定义的培养基。第一步是将在补充胎牛血清(FBS)的培养基中培养的细胞转移到您选择的含有相同量胎牛血清的化学定义培养基中。接下来的步骤包括逐步减少FBS的数量,同时监测细胞生长和活力,直到完全消除FBS。该方案已成功地将THP-1细胞适应于化学定义的培养基,具有与用FBS培养的细胞相似的最大特定生长速率。©2019 by John Wiley &基本方案:逐渐适应化学定义的介质替代方案:直接适应化学定义的介质支持方案1:确定细胞培养的最大特定生长速率支持方案2:细胞冷冻和解冻
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