Laboratoire des Sciences du Génie Chimique, UPR CNRS 6811, Nancy-Université, 2 avenue de la Forêt de Haye, 54505, Vandoeuvre-lès-Nancy Cedex, France.
Appl Biochem Biotechnol. 2010 Mar;160(6):1600-15. doi: 10.1007/s12010-009-8674-0. Epub 2009 Jun 9.
The paper proposes a rapid screening method for a first step improvement of an animal component-free medium dedicated to the growth of the anchorage-dependent Vero cell line. A new, rapid, and non-invasive technique is presented to specifically monitor cultures of adherent cells in 96-well plates. The operating conditions of an image analyzer are adapted to take into account the decrease of cell size when the attached cell density increases. An experimental design is carried out to assess the influence of ten component groups in the original medium. Two groups including protein extracts, growth factor, insulin, glucose, and pyruvate show significant positive effects. The groups with vitamins and molecules related to nitrogenous bases display a less pronounced influence. The mixture of amino acids, B(1) vitamin, magnesium sulfate, and sodium phosphate as well as the couple sodium citrate and ferric chloride lead to a downward trend. The screening results are proved to be scalable in stirred cultures with cells on microcarriers. An improved serum-free medium, with some component groups being removed or added, can be rapidly formulated to reach respectively similar or 1.6 times higher cell density than in the original medium. The results from this global approach could be helpful to further focus experiments on identified medium components.
本文提出了一种快速筛选方法，用于对专门用于锚定依赖型 Vero 细胞系生长的无动物成分培养基进行初步改进。本文提出了一种新的、快速的、非侵入性技术，用于专门监测 96 孔板中贴壁细胞的培养。图像分析仪的操作条件进行了调整，以考虑到当附着细胞密度增加时细胞体积的减小。通过实验设计来评估原始培养基中十个成分组的影响。两组包括蛋白质提取物、生长因子、胰岛素、葡萄糖和丙酮酸盐，对细胞生长有显著的积极影响。含维生素和含氮碱基相关分子的成分组显示出的影响不那么明显。包含氨基酸、B(1)维生素、硫酸镁和磷酸钠的混合物以及柠檬酸钠和氯化铁的组合则呈现下降趋势。在搅拌培养中用微载体培养细胞的筛选结果证明是可扩展的。可以快速配制去除或添加某些成分组的改良无血清培养基，以分别达到与原始培养基相似或 1.6 倍的细胞密度。这种全局方法的结果有助于进一步将实验集中在确定的培养基成分上。
