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| Isolation method | Isolation principle | Isolation technique |
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| Enzymatic [11, 188] | Digestion of the tissue extracellular components by proteolytic enzymes | (1) Use of proteolytic enzymes such as collagenase and trypsin to digest the extracellular matrix.
(2) After the extracellular matrix has dissolved, the released cells are seeded into culture dishes in growth medium. |
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| Explant culture [11, 195] | Cell surface charge and adhesion to plastic surfaces | (1) The tissue is rinsed to remove blood cells.
(2) The tissue is cut into smaller pieces of no more than a few millimeters in length.
(3) The pieces are placed in culture dishes or flasks with growth medium.
(4) Cells start to migrate out of tissue and adhere to the culture surface, and after several days, the tissue pieces can be removed. |
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| Density-gradient centrifugation methods [567] | Cell size and/or density | (1) The sample is positioned on one or more layers having distinct densities, which are intermediate between those of the cells that are to be isolated and all other cells in the sample.
(2) After that, centrifugation of the sample at the appropriate speed fractionates it into distinct phases between the different density layers. |
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| Fluorescence-activated cell sorting (FACS) [181, 568] | Fluorescently labeled antibodies bind to surface or intracellular molecules | (1) Cells are labeled with a mixture of fluorescently conjugated antibodies.
(2) The labeled cells pass aligned one by one through a nozzle which vibrates to produce droplets containing individual cells at a defined distance from the nozzle.
(3) As the cells pass through the light source, a computer registers their individual light scatter and multiple fluorescent properties to detect cells that meet the preestablished criteria for selection.
(4) A mild electrical charge is used to charge the drop where wanted cells are present. When the charged droplets pass between the two electrically charged metal plates, it deflects into a different collection tube. |
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| Magnetic-activated cell sorting (MACS) [181, 567] | Magnetically labeled antibodies bind to surface molecules | (1) Cells are labeled with antibodies conjugated to biodegradable iron-based nanobeads.
(2) The labeled cells pass through a strong magnetic field.
(3) Cells conjugated with magnetic particles stay on the column, while nonconjugated cells pass though. |
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