Bioprozess- und Bioverfahrenstechnik, Technische Universität Hamburg-Harburg, Denickestrasse 15, D-21071, Hamburg, Germany.
Cytotechnology. 1999 Jul;30(1-3):95-106. doi: 10.1023/A:1008055702079.
Apoptosis is an active, genetically determined death mechanism which can be induced by a wide range of physiological factors and by mild stress. It is the predominant form of cell death during the production of antibodies from murine hybridoma cell lines. A number of studies have now demonstrated that the suppression of this death pathway, by means of over-expression of survival genes such as bcl-2, results in improved cellular robustness and antibody productivity during batch culture. In the present study, the influence of bcl-2 expression on hybridoma productivity in two high density perfusion bioreactor systems was investigated. In the first system, a fixed-bed reactor, the DNA content in the spent medium was 25% higher in the control (TB/C3-pEF) culture than that found in the bcl-2 transfected (TB/C3-bcl2) cultures at all perfusion rates. This is indicative of a higher level of cell death in the control cell line. The average antibody concentration for the TB/C3-pEF cell line was 14.9 mg L-1 at perfusion rates of 2.6 and 5.2 d-1. However, for the TB/C3-bcl2 cell line it was 33 mg L-1 at dilution rates of 2 and 4 d-1. A substantial increase in antibody concentration was also found in the Integra Tecnomouse hollow fibre reactor. The antibody titre in the TB/C3-bcl2 cassette was nearly 100% higher than that in the TB/C3-pEF cassette during the cultivation period which lasted 6 weeks. Clearly, these results demonstrate the positive impact of bcl-2 over-expression on production of antibody in hybridoma perfusion cultures.
细胞凋亡是一种主动的、由基因决定的死亡机制，可被广泛的生理因素和轻度应激诱导。它是鼠杂交瘤细胞系产生抗体过程中主要的细胞死亡形式。现在有许多研究表明，通过过度表达生存基因（如 bcl-2）抑制这种死亡途径，可在批式培养过程中提高细胞的稳健性和抗体的生产力。在本研究中，研究了 bcl-2 表达对两种高密度灌注生物反应器系统中杂交瘤生产力的影响。在第一个固定床反应器中，在所有灌注率下，对照（TB/C3-pEF）培养物中消耗培养基中的 DNA 含量比 bcl-2 转染（TB/C3-bcl2）培养物中的高 25%。这表明对照细胞系中的细胞死亡水平更高。TB/C3-pEF 细胞系在灌注率为 2.6 和 5.2 d-1 时的平均抗体浓度为 14.9 mg L-1。然而，对于 TB/C3-bcl2 细胞系，在稀释率为 2 和 4 d-1 时，其平均抗体浓度为 33 mg L-1。在 Integra Tecnomouse 中空纤维反应器中也发现了抗体浓度的大幅增加。在培养期间，TB/C3-bcl2 盒的抗体滴度比 TB/C3-pEF 盒高近 100%，该培养期持续了 6 周。显然，这些结果表明 bcl-2 过表达对杂交瘤灌注培养中抗体生产的积极影响。
