Decellularized murine lung and brain provide matrix material for tissue culture studies

The extracellular matrix (ECM) is the natural support structure that surrounds cells in tissues, regulating how cells grow, move, and interact with their environment. ECM can also influence the phenotype and metastatic ability of cancer cells across organs. To investigate the role of ECM in regulating cancer cell phenotype, we developed a decellularization protocol for murine lung and brain tissues, providing a matrix material for growing the triple-negative human breast cancer cell line, MDA-MB-231. Brain and lung tissues from age- and sex-matched C57BL/6 mice were decellularized, cryosectioned, and processed for H&E and Masson's Trichrome staining or used for culturing RFP-Vimentin MDA-MB-231 cells. Optimization of the protocol incubation times and temperatures, as well as the concentrations of detergent and DNase treatments, resulted in a significant reduction in protein and DNA content in the lungs and brains, accompanied by a loss of cellular and nuclear staining in both organs. Cells were cultured within the decellularized brain and lung matrices, showing apparent differences in morphology and organization compared to cells grown on glass slides alone. These data demonstrate the feasibility of using flash-frozen organs to create decellularized matrices suitable for cell growth and the study of changes in cellular behavior and phenotype.