Linderholm Angela L, Bhandari Manohar P, Borras Eva, Kwon Allison, Herve Flore M, McCartney Mitchell M, Harper Richart W, Kenyon Nicholas J, Davis Cristina E
Division of Pulmonary, Critical Care and Sleep Medicine, University of California, Davis, Sacramento, CA, USA.
Lung Center, University of California, Davis, Davis, CA, USA.
Anal Bioanal Chem. 2025 Mar;417(6):1165-1176. doi: 10.1007/s00216-024-05720-z. Epub 2024 Dec 26.
Metabolically active cells emit volatile organic compounds (VOCs) that can be used in real time to non-invasively monitor the health of cell cultures. We utilized these naturally occurring VOCs in an adapted culture method to detect differences in culturing Chinese hamster ovary (CHO) cells with and without Staphylococcus epidermidis and Aspergillus fumigatus contaminations. The VOC emissions from the cell cultures were extracted and measured from the culture flask headspace using polydimethylsiloxane (PDMS)-coated Twisters, which were subjected to thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS) analysis. In our initial time points of 1 and 2 h, we detected VOC signatures that differentiated the cultures earlier than traditional plating techniques or visualization methods. Partial least squares-discriminant analysis (PLS-DA) models were built to differentiate the analytes from the CHO cells and S. epidermidis- and A. fumigatus-inoculated CHO cultures. A total of 41 compounds with a variable importance in projection (VIP) score greater than 1 was obtained across the models. Similarly, based on the PLS regression analyses to predict the cell concentration of S. epidermidis (R = 0.891) and A. fumigatus (R = 0.375), 15 and 20 relevant compounds were putatively identified, respectively; two known compounds overlapped between the two microbes. Some of the compounds were unidentified and future studies will determine the relationship between the VOCs and the metabolic changes in contaminated cultures.
代谢活跃的细胞会释放挥发性有机化合物（VOCs），这些化合物可用于实时、非侵入性地监测细胞培养物的健康状况。我们在一种改良的培养方法中利用这些天然存在的VOCs，来检测接种和未接种表皮葡萄球菌及烟曲霉的中国仓鼠卵巢（CHO）细胞培养物之间的差异。使用涂有聚二甲基硅氧烷（PDMS）的Twisters从培养瓶顶空提取并测量细胞培养物释放的VOCs，然后对其进行热解吸-气相色谱-质谱（TD-GC-MS）分析。在最初的1小时和2小时时间点，我们检测到的VOC特征比传统的平板接种技术或可视化方法能更早地区分不同的培养物。构建了偏最小二乘判别分析（PLS-DA）模型，以区分来自CHO细胞以及接种了表皮葡萄球菌和烟曲霉的CHO培养物中的分析物。在所有模型中，共获得了41种投影变量重要性（VIP）得分大于1的化合物。同样，基于PLS回归分析来预测表皮葡萄球菌（R = 0.891）和烟曲霉（R = 0.375）的细胞浓度，分别推定鉴定出15种和20种相关化合物；两种微生物之间有两种已知化合物重叠。其中一些化合物尚未鉴定出来，未来的研究将确定VOCs与受污染培养物中代谢变化之间的关系。
