The impact of altered matriptase activity on cell viability and spheroid growth in 2 and 3D cell cultures

Abstract

Anticancer drug research requires suitable cell model systems to accurately screen and develop new treatment options capable of acting against tumour growth and metastasis. Currently the most suitable models are live animals, however nowadays, due to animal welfare and protection concerns, other potential replacement methods are being explored to minimise the use of live animals in pharmaceutical research. In 3D or multilayer in vitro cell cultures cells can produce their own extracellular matrix, which can influence the uptake and transport of drugs, thus better mimicking the in vivo conditions than the widely used monolayer cell culture systems. Our goals in the experiments were to characterise the cells' viability using MTS assay, and the size and cell numbers of the spheroids. We used non-specific AEBSF and specific matriptase inhibitors such as MI-432, MI-453, MI-460 and MI-463 at a concentration of 50 µM, and a matriptase inducer, suramin, at concentrations of 50, 100 and 200 µM. During our research, we used three different cell lines- a porcine neonatal non-tumorigenic jejunal cell line, IPEC-J2, a murine colonic adenocarcinoma cell line, C26 and a human colon adenocarcinoma cell line, HT-29.