Optimisation of methods for isolation of primary cancer cells from breast cancer tumour

Abstract

Abstract Primary cell lines are an invaluable tool to explore cancer biology and molecular processes and investigate novel treatments. Despite their numerous advantages, primary cultures are laborious to obtain and maintain in culture thus rarely used in the laboratory. In this paper, we tested various methods for isolating primary breast cancer cultures that differed in enzymatic composition, incubation times and other methodologies. Finally, we optimised a method (Method 5) that gave rise to primary cell culture BC160. This approach combines mechanical disaggregation with enzymatic digestion by collagenase IV and hyaluronidase, followed by centrifugation. The cell line obtained by this method has a unique set of features with a strong influence of extracellular matrix remodelling genes. Primary cell lines have a high potential to become a model of choice, hence it is essential to elaborate and categorise isolation methods, develop approaches to separate heterogeneous cultures and investigate factors influencing the establishment of primary cell lines.