Presentation: 2025 ND EPSCoR Annual conference
October 21, 2025, NDSU Memorial Union, Fargo, North Dakota
Decellularized Murine Lungs and Brains Provide Matrix Material for Tissue Culture Studies
Natalia
Focsa
Lab Technician
University of North Dakota
Co-authors: Angela M. Floden, Senior Research Specialist, Biomedical Science Department, University of North Dakota, Chen Cheng, Post Doc Research Fellow Lab, Biomedical Science Department, University of North Dakota, Colin K Combs, Department Chair, Sr Assoc Dean/Assoc AVPR, Research Affairs, University of North Dakota
Session
Poster number: 35
Ballroom
The extracellular matrix (ECM) is the natural support structure that surrounds cells in tissues where it regulates how cells grow, move, and interact with their environment. ECM can also influence the phenotype and ability of cancer cells to metastasize across organs. To begin studying the role of ECM in regulating cancer cell phenotype, we developed a decellularization protocol for murine lung and brain tissues to provide a matrix material for growing the triple negative human breast cancer cell line, MDA-MB-231. Brain and lungs tissues from age- and sex-matched C57BL/6 mice were aseptically collected and immediately flash frozen in liquid nitrogen. The tissues were subsequently thawed in ice-cold sterile distilled water and protein and nucleic acid levels reduced by a series of incubations in detergents and DNase. The decellularized tissues were cryosectioned and processed for H&E and Masson’s Trichrome staining or used for culturing RFP-Vimentin MDA-MB-231 cells. The optimization of the protocol for incubation times and temperatures as well as the concentrations of detergent and DNAse resulted in a significant reduction in both protein and DNA content in lungs and brains. These observations were further supported by loss of cellular and nuclear staining in both organs while the tissue integrity remained intact. Cells were cultured on the sectioned tissue for four days and they populated within the decellularized brain and lung matrices with clear differences in morphology and organization compared to cells grown on the glass slides alone. These data demonstrate the feasibility of using flash frozen organs for creating decellularized matrices that are appropriate for cell growth and to study changes in cellular behavior and phenotype.