Schmale Kathrin, Rademacher Thomas, Fischer Rainer, Hellwig Stephan
Institut für Biologie VII (Molekulare Biotechnologie), RWTH Aachen University, Germany.
J Biotechnol. 2006 Jun 25;124(1):302-11. doi: 10.1016/j.jbiotec.2006.01.012. Epub 2006 Mar 10.
Plants have been established as an useful production system for commercially relevant proteins. Plant cell cultures show certain advantages compared to field-grown plants. However, one critical drawback for the long-term use of recombinant plant cell cultures is the instability of cell cultures concerning genetic background and productivity when maintained by subculturing. The ability to store recombinant cell lines stably by cryopreservation allows to maintain an efficient and stable production system. In this work, we describe the development of a cryopreservation protocol for a transgenic BY-2 cell culture expressing human serum albumin. In 1.75-L stirred-tank bioreactors growth and production kinetics of the transgenic cell line were compared after cryopreservation to the hitherto performed maintenance by subculturing. Growth and productivity of the cryopreserved cell culture remained stable after freezing for one week. Here, we show that we developed an efficient method which allows the storage of transgenic plant cell cultures, an important requirement for industrial processes.
植物已被确立为生产具有商业价值蛋白质的有用生产系统。与田间种植的植物相比，植物细胞培养显示出某些优势。然而，重组植物细胞培养长期使用的一个关键缺点是，通过传代培养维持时，细胞培养物在遗传背景和生产力方面的不稳定性。通过冷冻保存稳定储存重组细胞系的能力有助于维持高效稳定的生产系统。在这项工作中，我们描述了一种用于表达人血清白蛋白的转基因BY-2细胞培养物的冷冻保存方案的开发。在1.75升搅拌罐生物反应器中，将冷冻保存后的转基因细胞系的生长和生产动力学与迄今通过传代培养进行的维持情况进行了比较。冷冻一周后，冷冻保存的细胞培养物的生长和生产力保持稳定。在这里，我们表明我们开发了一种有效的方法，该方法允许储存转基因植物细胞培养物，这是工业生产过程的一项重要要求。
