Institute of Chemical Engineering, Swiss Federal Institute of Technology (EPFL), CH-1015, Lausanne, Switzerland.
Cytotechnology. 2001 Oct;37(2):65-73. doi: 10.1023/A:1019956013627.
A balanced supplementation method was applied to develop a serum and protein- free medium supporting hybridoma cell batch culture. The aim was to improve systematically the initial formulation of the medium to prevent limitations due to unbalanced concentrations of vitamins and amino acids. In a first step, supplementation of the basal formulation with 13 amino acids, led to an increase of the specific IgA production rate from 0.60 to 1.07 pg cell(-1) h(-1). The specific growth rate remained unchanged, but the supplementation enabled maintenance of high cell viability during the stationary phase of batch cultures for some 70 h. Since IgA production was not growth- related, this resulted in an approximately4-fold increase in the final IgA concentration, from 26.6 to 100.2 mgl(-1). In a second step, the liposoluble vitamins E and K(3) were added to the medium formulation. Although this induced a slightly higher maximal cell concentration, it was followed by a sharp decline phase with the specific IgA production rate falling to 0.47 pg cell(-1) h(-1). However, by applying a second cycle of balanced supplementation with amino acids this decline phase could be reduced and a high cell viability maintained for over 300 h of culture. In this vitamin- and amino acid- supplemented medium, the specific IgA production rate reached a value of 1.10 pg cell(-1)h(-1) with a final IgA concentration of 129.8 mgl(-1). The latter represents an increase of approximately5-fold compared to the non- supplemented basal medium.
采用平衡补充法开发了一种支持杂交瘤细胞分批培养的无血清和无蛋白培养基。目的是系统地改进初始培养基配方，以防止由于维生素和氨基酸浓度不平衡而导致的限制。在第一步中，用 13 种氨基酸补充基础配方，使特异性 IgA 生产速率从 0.60 增加到 1.07 pg 细胞(-1) h(-1)。特异性生长率保持不变，但补充剂使一些 70 小时的分批培养静止期的高细胞活力得以维持。由于 IgA 生产与生长无关，因此最终 IgA 浓度从 26.6 增加到 100.2 mgl(-1)，大约增加了 4 倍。在第二步中，将脂溶性维生素 E 和 K(3)添加到培养基配方中。尽管这诱导了稍高的最大细胞浓度，但随后是一个急剧下降阶段，特异性 IgA 生产速率下降到 0.47 pg 细胞(-1) h(-1)。然而，通过应用第二轮氨基酸平衡补充，可以减少下降阶段并保持超过 300 小时的高细胞活力。在这种添加维生素和氨基酸的培养基中，特异性 IgA 生产速率达到 1.10 pg 细胞(-1) h(-1)，最终 IgA 浓度为 129.8 mgl(-1)。与未补充的基础培养基相比，后者增加了约 5 倍。
