Marigliani Bianca, Balottin Luciene Bottentuit López, Augusto Elisabeth de Fatima Pires
PRONAMETRO, National Institute of Metrology, Quality and Technology (INMETRO), Duque de Caxias, Rio de Janeiro, Brazil.
Laboratory of Tissue Bioengineering, Directory of Metrology Applied to Life Sciences, National Institute of Metrology, Quality and Technology (INMETRO), Duque de Caxias, Rio de Janeiro, Brazil.
Curr Protoc Toxicol. 2019 Dec;82(1):e88. doi: 10.1002/cptx.88.
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.
为了规避在细胞培养中使用动物血清所涉及的伦理、技术和经济方面的弊端，可以使哺乳动物细胞适应无血清培养基。如今，有几种无血清配方可供选择，包括完全无动物源和化学成分明确的培养基，以及不同的适应技术。本文重点介绍哺乳动物悬浮细胞培养逐步适应化学成分明确的培养基的过程。第一步是将在补充有胎牛血清（FBS）的培养基中培养的细胞转移到你选择的含有相同量FBS的化学成分明确的培养基中。接下来的步骤包括逐步减少FBS的量，同时监测细胞生长和活力，直至完全去除FBS。该方案已成功用于使THP-1细胞适应化学成分明确的培养基，其最大比生长速率与用FBS培养的细胞相似。© 2019约翰威立国际出版公司。基本方案：逐步适应化学成分明确的培养基 替代方案：直接适应化学成分明确的培养基 支持方案1：测定细胞培养物的最大比生长速率 支持方案2：细胞冻存和解冻。
