Isolation and maintenance of commonly used primary cells for cell death research

This protocol provides detailed methods for the preparation, culture, and maintenance of two essential primary cell lines used in cell death studies: mouse embryonic fibroblasts (MEFs) and bone marrow-derived macrophages (BMDMs). These primary cells serve as valuable models for investigating molecular mechanisms of cell death pathways due to their physiological relevance compared to immortalized cell lines. The protocol covers isolation techniques from mouse embryos (for MEFs) and bone marrow (for BMDMs), differentiation protocols, culture conditions, cryopreservation methods. This comprehensive guide enables researchers to establish reliable primary cell culture systems for apoptosis, necroptosis, pyroptosis, and other cell death-related studies.