Radford K, Niloperbowo W, Reid S, Greenfield P F
Department of Chemical Engineering, University of Queensland, Brisbane.
Cytotechnology. 1991 May;6(1):65-78. doi: 10.1007/BF00353704.
A general weaning procedure is described which allowed a range of hybridomas to be weaned readily off serum without loss of antibody production. Initial work was carried out with one cell line only (SPO1 cells) and one serum substitute containing a final protein concentration of 40 mg l-1. The SPO1 cells were first adapted to a range of readily available basal media and then weaned off serum by a range of protocols. From this work an optimal weaning protocol and basal medium for weaning were determined. These were then used to wean the SPO1 cells and two other cell lines off serum with a second, protein free, serum substitute with varying concentrations of defined proteins added. All three cell lines investigated were readily weaned off serum by this protocol at protein concentrations as low as 1 mg l-1. No loss of antibody production was observed with any of the cell lines. The weaning procedure outlined in both simple and rapid and has been successfully adopted in our laboratory by relatively inexperienced cell culture technicians.
本文描述了一种通用的细胞系血清撤换方法，该方法可使多种杂交瘤细胞轻松实现血清撤换，且不会导致抗体产生量的损失。最初的实验仅使用了一种细胞系（SPO1细胞）和一种最终蛋白质浓度为40 mg l-1的血清替代品。首先使SPO1细胞适应一系列易于获取的基础培养基，然后通过一系列方案使其脱离血清培养。基于此项工作，确定了最佳的血清撤换方案和基础培养基。随后，使用这些方案，采用第二种不含蛋白质但添加了不同浓度特定蛋白质的血清替代品，使SPO1细胞和另外两种细胞系脱离血清培养。研究的所有三种细胞系均可通过该方案在低至1 mg l-1的蛋白质浓度下轻松实现血清撤换，且未观察到任何细胞系的抗体产生量有所损失。所述血清撤换方法既简单又快速，相对缺乏经验的细胞培养技术人员已在我们实验室成功采用该方法。
