A technique for monitoring mammalian cell growth and inhibition in situ via Fourier transform infrared spectroscopy.
A technique for monitoring mammalian cell growth and inhibition in situ via Fourier transform infrared spectroscopy.
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一种通过傅里叶变换红外光谱原位监测哺乳动物细胞生长和抑制的技术。
DOI：
10.1016/0003-2697(88)90040-1
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发表时间：
1988
影响因子：
2.9
通讯作者：
Chang,MJ
中科院分区：
文献类型：
--
作者：
Hutson,TB;Mitchell,ML;Keller,JT;Long,DJ;Chang,MJ
中国仓鼠卵巢细胞的单一培养物在锗衰减全反射 (ATR) 晶体上生长，并通过 ATR/傅里叶变换红外 (FT-IR) 光谱在原位连续监测约 60 小时。将细胞接种到专门设计的流动池中，该流动池控制生理条件、流速以及生长培养基或代谢抑制剂的添加。以 20 分钟的间隔采集红外光谱，直至形成汇合的单层。光谱中明显有几个强谱带，通常可归因于细胞成分的分子特征。细胞生长动力学被测量为红外带强度随时间变化的函数，并在达到汇合时表现出正常的滞后期、对数生长和稳定期。生长细胞的光谱（标准化为光谱区域 1800-1000 cm−1 下的面积）从 60 小时汇合细胞的参考光谱中减去。差异光谱显示汇合细胞和早期生长阶段细胞之间观察到的最大差异。差异可能反映细胞形态变化、生化活性以及 ATR 晶体暴露于大量培养基的程度。活体中国仓鼠卵巢细胞的 ATR FT-IR 光谱也被用于毒理学研究，以监测羟基脲（一种 DNA 合成抑制剂）的影响。与对照培养物相比，在处理过程中羟基脲处理的细胞的细胞生长曲线中观察到生长延迟。
A single culture of Chinese hamster ovary cells was grown on germanium attenuated total reflectance (ATR) crystals and continuously monitored in situ via ATR/Fourier transform infrared (FT-IR) spectroscopy for ∼60 h. The cells were seeded into a specially designed flow cell which controlled physiological conditions, flow rate, and addition of growth medium or metabolic inhibitors. Infrared spectra were taken at 20-min intervals until a confluent monolayer was formed. Several strong bands are evident in the spectra which can be generally ascribed to molecular features of cellular components. Cell growth kinetics were measured as a function of infrared band intensity over time and exhibited the normal lag phase, logarithmic growth, and stationary phase on reaching confluence. Spectra of growing cells, normalized to the area under the spectral region 1800-1000 cm−1, were subtracted from reference spectra of confluent cells at 60 h. Difference spectra showed that the largest differences were observed between confluent cells and cells in early growth stages. Differences may reflect cell morphological changes, biochemical activity, and degree of ATR crystal exposure to the bulk medium. ATR FT-IR spectroscopy of living Chinese hamster ovary cells was also used in a toxicological study to monitor the effects of hydroxyurea, an inhibitor of DNA synthesis. Delayed growth was observed in the cell growth curve of the hydroxyurea-treated cells during the course of treatment as compared to the control culture.