Colorectal cancer is one of the most commonly diagnosed and poorly responding to chemotherapy types of cancer, which emphasizes the importance of personalized approach to treatment selection. Short-term primary cell cultures established from patients’ tumors represent a valuable model for testing drug response. In this study, we developed protocols for generation of the short-term primary cell cultures from colorectal cancer tissue and assessment their chemosensitivity using MTT test. Additionally, we showed the possibility of metabolic analysis of patient-derived cancer cells using fluorescence lifetime imaging (FLIM) of autofluorescent cofactor NAD(P)H. Since FLIM of NAD(P)H demonstrates the potential to detect early responses to cancer treatment, we assume that this method, alone or in combination with MTT assay, can be used for choosing the optimal chemotherapy for patients.
Cirrhosis is defined as the histological development of regenerative nodules surrounded by fibrous bands in response to chronic liver injury that leads to portal hypertension and end stage liver disease. Conventional techniques are insufficient to precisely describe the internal structure, heterogeneous cell populations and the dynamics of biological processes of the diseased liver. Currently, multiphoton microscopy with fluorescence lifetime imaging is actively introduced to biomedical research. This technic is extremely informative and non-destructive that allows studying of a large number of processes occurring inside cells and tissues, analyzing molecular cellular composition, as well as evaluating the state of connective tissue fibers due to their ability to generate a second optical harmonic. In this study we investigated metabolic changes and collagen fibers formation in the rat liver with induced cirrhosis based on the fluorescence of the metabolic co-factors (NAD(P)H, FAD) and a second harmonic signal by multiphoton microscopy with FLIM and SHG mode. Moreover we studied ex vivo liver samples of patients with cirrhosis. We presented a separate analysis of NADH and NADPH to estimate the contribution of energy metabolism and lipogenesis in the metabolic changes. The data can be used to develop new criteria for the identification of hepatic pathology at the level of hepatocyte changes directed to personalized medicine in the future.
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