Protein Science Group, University of Kent, Canterbury, Kent, CT2 7NJ, UK.
Cytotechnology. 2007 Apr;53(1-3):47-53. doi: 10.1007/s10616-007-9048-5. Epub 2007 Feb 23.
In recent years there have been a number of reports that suggest the sub-physiological (<37 degrees C) temperature in vitro culturing of mammalian cells can result in enhanced heterologous protein production. Despite these reports, the mechanisms by which mammalian cells respond to such conditions are largely unknown. We therefore set out to use a model in vitro culture HeLa cell system to begin investigating the cold-shock response in mammalian cell systems. Sub-physiological temperature cultivation resulted in reduced growth and proliferation and a lower total cell protein content. Proteomic analysis confirmed that HeLa cells actively respond to sub-physiological temperature by up-regulating a number of proteins and immunoblot analysis confirmed that specific proteins are indeed up-regulated in a time and temperature dependent manner. Additional work is likely to improve our understanding of the cold-shock response in mammalian cells and identify candidate target proteins for cell engineering to further enhance heterologous protein production at sub-physiological temperatures.
近年来有许多报道表明,在亚生理温度(<37°C)下体外培养哺乳动物细胞可以提高异源蛋白的产量。尽管有这些报道,但哺乳动物细胞对这种条件的反应机制在很大程度上尚不清楚。因此,我们着手使用体外培养 HeLa 细胞系的模型开始研究哺乳动物细胞系统中的冷休克反应。亚生理温度培养导致细胞生长和增殖减少,总细胞蛋白含量降低。蛋白质组学分析证实,HeLa 细胞通过上调许多蛋白质来积极响应亚生理温度,免疫印迹分析证实特定蛋白质确实以时间和温度依赖性方式上调。进一步的工作可能会提高我们对哺乳动物细胞冷休克反应的理解,并确定用于细胞工程的候选靶蛋白,以进一步提高亚生理温度下异源蛋白的产量。