Tumor cell invasion into Matrigel: Optimized protocol for RNA extraction

Abstract

3D models are increasingly used to study mechanisms driving tumor progression and mimicking in vitro processes such as invasion and migration. However, there is a need to establish more protocols based on 3D culture systems that allow for downstream molecular biology investigations. Materials & methods: Here we present a method for optimal RNA extraction from highly aggressive primary glioma cells invading into Matrigel. The method has been established by comparing previously reported protocols, available commercial kits and optimizing specific steps for matrix dissociation, RNA separation and purification. Results and conclusion: The protocol allows RNA extraction from cells embedded into Matrigel, with optimal yield, purity and integrity suitable for subsequent sequencing analysis of both high and low molecular weight RNA. METHOD SUMMARY An optimized method for RNA extraction fromhighly invasive glioma cells embedded intoMatrigel was achieved by using TRIzol, used formatrix dissociation, cell lysis and RNA separation, in combination with a specific commercial kit used to perform the DNAse treatment and optimal RNA purification on column. The method ensures optimal yield, good purity and integrity for analysis of high and low molecular weight RNA molecules.