Lifanov Dmitry, Zorigt Dulamsuren, Shabalina Evgenya, Khalil Abdullah, Gorbunov Konstantin, Petersen Elena
Institute of Future Biophysics, Institutskiy per. 9, Dolgoprudny, Moscow Oblast, Moscow, Russia.
Scientific Research Institute for Systems Biology and Medicine, Nauchny prd. 18, Moscow, Russia.
BMC Mol Cell Biol. 2025 Jan 28;26(1):7. doi: 10.1186/s12860-025-00532-0.
This paper describes a method for determining the cytotoxicity of chemical compounds based on the detection of fluorescent proteins-in this case, green fluorescent protein (GFP) and red fluorescent protein (RFP), which are released into the medium from dead cells. This method is similar in principle to the lactate dehydrogenase test (LDH test), but it does not require a reaction with a chromogenic substrate. This method also makes it possible to independently determine the viability of different lines when used in cocultures. Experiments were performed on a classical monolayer, spheroids and 3D cultures in alginate hydrogel. Capecitabine was used as a model cytotoxic agent. We included liver cells (Huh7) in a coculture model and determined changes in the cytotoxicity levels of capecitabine against NCI-H1299 cells. The experimental part also found that there were differences in sensitivity to capecitabine depending on the type of 3D cultures used.
本文描述了一种基于检测荧光蛋白来测定化合物细胞毒性的方法——在这种情况下，是检测从死细胞释放到培养基中的绿色荧光蛋白（GFP）和红色荧光蛋白（RFP）。该方法在原理上与乳酸脱氢酶测试（LDH测试）相似，但它不需要与显色底物发生反应。当用于共培养时，该方法还能够独立测定不同细胞系的活力。实验在经典单层培养、球体培养以及藻酸盐水凝胶中的三维培养上进行。使用卡培他滨作为细胞毒性模型药物。我们在共培养模型中纳入了肝细胞（Huh7），并测定了卡培他滨对NCI-H1299细胞的细胞毒性水平变化。实验部分还发现，根据所使用的三维培养类型，对卡培他滨的敏感性存在差异。
