Types of cell death and apoptotic stages in Chinese Hamster Ovary cells distinguished by Raman spectroscopy
Types of cell death and apoptotic stages in Chinese Hamster Ovary cells distinguished by Raman spectroscopy
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DOI：
10.1002/bit.26476
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发表时间：
2018-02-01
影响因子：
3.8
通讯作者：
Piret, James M.
中科院分区：
文献类型：
--
作者：
Rangan, Shreyas;Kamal, Sepehr;Piret, James M.
细胞死亡是用于生产治疗性蛋白质的生物反应器中生产力损失的最终原因。我们研究了拉曼光谱检测中国卵巢（CHO）细胞的细胞死亡的发生和类型的能力-最广泛使用的细胞类型用于治疗性蛋白质生产。拉曼光谱用于比较凋亡、坏死、自噬和对照CHO细胞。在650-850 cm（-1）光谱范围内确定了几个特定的核酸、蛋白质和脂质相关标记带，这些标记带可区分经历不同细胞死亡模式的细胞;支持证据由全光谱数据的主成分分析（PCA）提供。除了比较细胞死亡的不同模式之外，还将正常细胞与在凋亡的几个阶段分选的细胞进行比较，以探索早期检测凋亡的潜力。通过拉曼光谱可以区分细胞凋亡的不同阶段，在早期阶段观察到核酸峰的多种变化，而脂质信号的增加是晚期细胞凋亡/继发性坏死的特征。
Cell death is the ultimate cause of productivity loss in bioreactors that are used to produce therapeutic proteins. We investigated the ability of Raman spectroscopy to detect the onset and types of cell death for Chinese Hamster Ovary (CHO) cells-the most widely used cell type for therapeutic protein production. Raman spectroscopy was used to compare apoptotic, necrotic, autophagic, and control CHO cells. Several specific nucleic acid-, protein-, and lipid-associated marker bands within the 650-850 cm(-1) spectral region were identified that distinguished among cells undergoing different modes of cell death; supporting evidence was provided by principal component analysis (PCA) of the full spectral data. In addition to comparing the different modes of cell death, normal cells were compared to cells sorted at several stages of apoptosis, in order to explore the potential for early detection of apoptosis. Different stages of apoptosis could be distinguished via Raman spectroscopy, with multiple changes observed in nucleic acid peaks at early stages whereas an increase in lipid signals was a feature of late apoptosis/secondary necrosis.