Department of Pediatrics, Medical University of Vienna, Wahringer Gurtel 18, 1090 Vienna, Austria.
J Proteome Res. 2009 Nov;8(11):5285-95. doi: 10.1021/pr900630s.
Stability of cell lines is the prerequisite for all in vitro research, but literature on the stability of protein expression over passages is limited. Determination of specific stability markers, karyotyping, and morphology may not provide full information on this subject. It was the aim of the study to test protein level fluctuations in a human amniotic fluid stem cell line from passages 5, 7, 11, and 25. While karyotype, cell cycle, apoptosis rate, and 10 markers for characterization of the cell line remained unchanged (carried out at passages 5 and 25), cell volume was increased at passage 25. Significant protein fluctuations were observed for signaling, antioxidant, guidance cue, proteasomal, connective tissue, cytoskeleton proteins, chaperones, a chloride channel, and prothymosin at passages 5, 7, 11, and 25. Herein, the use of this gel-based proteomic screen, checking protein stability for the characterization of cell lines in addition to corresponding published markers, is proposed, in particular when experiments are run over several passages.
细胞系的稳定性是所有体外研究的前提,但关于蛋白质表达在传代过程中的稳定性的文献有限。确定特定的稳定性标志物、核型分析和形态学可能无法提供关于这个主题的完整信息。本研究的目的是检测人羊水干细胞系在第 5、7、11 和 25 代时的蛋白质水平波动。虽然核型、细胞周期、凋亡率和 10 个用于鉴定细胞系的标志物在第 5 和 25 代时保持不变(在第 5 和 25 代进行),但在第 25 代时细胞体积增加。在第 5、7、11 和 25 代时,观察到信号转导、抗氧化、导向线索、蛋白酶体、结缔组织、细胞骨架蛋白、伴侣蛋白、氯离子通道和胸腺素前体等蛋白质出现显著波动。在此,提出了使用这种基于凝胶的蛋白质组学筛选方法,除了相应的已发表标志物外,还可以检查蛋白质稳定性来鉴定细胞系,特别是在进行多个传代实验时。