Animal Cell Separation and Subcellular Fractionation

Abstract Cell populations isolated from tissues or from blood are remarkably complex containing many different types that can be dissociated and separated based on cell phenotype. Sorting and separation strategies differ widely as do the resulting cell populations in terms of both parameters upon which the sort is based and sample purity. Separation strategies can be simple or complex and based on culture characteristics or gene expression profiles. Each cell population itself contains many functionally different subcellular compartments that can also be selectively fractionated into subcellular components. Fractionation strategies must be tailored to provide the highest quality enrichment of the fraction of cells, organelles or complexes sought and after careful assessment of the target cell or molecule and the uses to which the final enriched fraction will be put. Key Concepts: Complex populations of cells from tissues or blood can be liberated and then sorted or separated for culture or subcellular fractionation. Sorted cell populations can be fractionated into organelle compartments based on differential use of dilute detergents and differential centrifugation. Sorted cell populations can be fractionated into functional subcellular compartments based on affinity of different molecules for the membrane and fibre systems within cells. Careful evaluation of the purity and authenticity of each cell and molecular fraction is required before use or biological evaluation.