Isolation and Purification of Various Mammalian Cells: Single Cell Isolation

Abstract

A typical adult mammalian body consists of 30–40 trillion cells. Mammalian cells differ in their shape, size, morphology, and other structural and functional characteristics. These characteristics of mammalian cells can be the basis for the isolation of a particular cell type from any tissue or organ. Except for blood cells, various other mammalian cells are firmly adjoined together by extracellular matrix (ECM) proteins to form tissues and organs. To isolate and enrich the greater number of cells, their dislodging or enzymatic dissociation from the original parent tissues or organs is highly essential. Following dissociation, the isolation and purification of mammalian cells may be based on the following procedures: (1) Cell isolation is based on cell surface charge and adhesion. (2) Cell isolation based on cell size and density (density gradient centrifugation, filtration, sedimentation). (3) Cell isolation based on cell morphology and physiology (selective culture medium, laser capture microdissection system). (4) Cell isolation based on cell surface markers (fluorescence-activated cell sorting, magnetic separation, cell purification using complement depletion, purification methods involving DNA sequences, other technology: mitochondrial dye and aptamer technology). (5) Cell isolation based on a combination of the above techniques (immuno-density-assisted separation: erythrocyte Rosetting immuno-laser capture microdissection system, microfluidics-based cell isolation, hydrodynamic cell sorting, acoustic cell sorting, electrophoretic sorting of cells). In this chapter, besides describing all the above-mentioned isolation techniques, toward the end, various single-cell isolation techniques and the tools for single-cell analysis such as genomics, transcriptomics, and proteomics are discussed.