Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, Krakow, Poland.
Department of Cell Biology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
J Biophotonics. 2019 Feb;12(2):e201800152. doi: 10.1002/jbio.201800152. Epub 2018 Oct 25.
Confocal Raman imaging combined with fluorescence-activated cell sorting was used for in vitro studies of cell cultures to look at biochemical differences between the cells in different cell phases. To answer the question what is the impact of the cell cycle phase on discrimination of pathological cells, the combination of several factors was checked: a confluency of cell culture, the cell cycle dynamics and development of pathology. Confluency of 70% and 100% results in significant phenotypic cell changes that can be also diverse for different batches. In 100% confluency cultures, cells from various phases become phenotypically very similar and their recognition based on Raman spectra is not possible. For lower confluency, spectroscopic differences can be found between cell cycle phases (G /G , S and G /M) for control cells and cells incubated with tumor necrosis factor alpha (TNF-α), but when the mycotoxin cytochalasin B is used the Raman signatures of cell phases are not separable. Generally, this work shows that heterogeneity between control and inflamed cells can be bigger than heterogeneity between cell cycle phases, but it is related to several factors, and not always can be treated as a rule.
共聚焦拉曼成像与荧光激活细胞分选相结合,用于体外细胞培养研究,以观察不同细胞期细胞之间的生化差异。为了回答细胞周期阶段对病理性细胞识别的影响这个问题,检查了几个因素的组合:细胞培养的汇合度、细胞周期动力学和病理学的发展。70%和 100%的汇合度导致显著的表型细胞变化,对于不同批次的细胞也可能是多样化的。在 100%汇合度的培养物中,来自不同阶段的细胞在表型上变得非常相似,并且基于拉曼光谱对它们的识别是不可能的。对于较低的汇合度,可以在对照细胞和用肿瘤坏死因子-α(TNF-α)孵育的细胞之间的细胞周期阶段(G1/G0、S 和 G2/M)之间找到光谱差异,但当使用真菌毒素细胞松弛素 B 时,细胞周期阶段的拉曼特征不可分离。一般来说,这项工作表明,对照细胞和炎症细胞之间的异质性可能比细胞周期阶段之间的异质性更大,但这与几个因素有关,并不总是可以作为一个规则来对待。