Hammoudeh Sarah Musa, Hammoudeh Arabella Musa, Hamoudi Rifat
Sharjah Institute for Medical Research, College of Medicine, University of Sharjah, 27272, Sharjah, UAE.
Department of Clinical Sciences, College of Medicine, University of Sharjah, Sharjah, UAE.
Histochem Cell Biol. 2019 Jul;152(1):75-84. doi: 10.1007/s00418-019-01775-7. Epub 2019 Feb 16.
One of the main aspects investigated in potential therapeutic compounds is their effect on cells viability and proliferative ability. Although various methods have been developed to investigate these aspects, these methods present with shortcomings in terms of either cost, availability, accuracy, precision, or throughput. This study describes a simple, economic, reproducible, and high-throughput assay to quantify cell death and proliferation. In this assay, adherent cells are fixed, stained with trypan blue, and measured for trypan blue internalization using a spectrophotometric absorbance plate reader. Corresponding cell counts to the absorbance measurements are extrapolated from a standard curve. This assay was used to measure the effect of dimethyl sulfoxide (DMSO) on the viability of breast and lung cancer cells. Decrease in cell count associated with the increase in DMSO percentage and exposure time. The assay's results closely correlated with the conventional trypan blue exclusion assay (Pearson correlation coefficient (r) > 0.99; p < 0.0001), but with higher precision. The assay developed in this study can be used for various applications such as optimization, cell treatment investigations, proliferation, and cytotoxicity studies.
潜在治疗化合物研究的主要方面之一是它们对细胞活力和增殖能力的影响。尽管已经开发出各种方法来研究这些方面,但这些方法在成本、可用性、准确性、精密度或通量方面都存在不足。本研究描述了一种简单、经济、可重复且高通量的测定方法,用于量化细胞死亡和增殖。在该测定中,贴壁细胞被固定,用台盼蓝染色,并使用分光光度吸光板读数器测量台盼蓝内化情况。从标准曲线外推与吸光度测量相对应的细胞计数。该测定方法用于测量二甲基亚砜(DMSO)对乳腺癌和肺癌细胞活力的影响。细胞计数的减少与DMSO百分比和暴露时间的增加相关。该测定结果与传统的台盼蓝排斥试验密切相关(皮尔逊相关系数(r)>0.99;p<0.0001),但具有更高的精密度。本研究中开发的测定方法可用于各种应用,如优化、细胞治疗研究、增殖和细胞毒性研究。