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subculture

Subculture (passaging) refers to transferring cells grown in a culture vessel to a new vessel to maintain culture continuously.
In the case of adherent cells, the cell adhesion is dissociated using a cell dissociation solution to remove cells from the culture vessel and transfer to a new culture vessel. For suspension cells, collect the culture and transfer to a new culture vessel. Most of the culture medium is replaced with fresh medium, but depending on the cell type, a small amount of old medium may be added to a new culture vessel. Passaging is sometimes performed when cells have grown sufficiently, but it is also performed when some cells have come too much into contact. In subcultures where cells are growing adequately, it is desirable to keep the cell dilution constant to maintain the phenotype. For example, 3T3 cells are passaged at a density of 3x10⁵ cells/60 mm Petri dish and passaged every three days. Depending on each cell line, there is an appropriate passaging method for maintaining phenotype. Therefore, passaging should be performed after obtaining information from papers on the establishment of the cell lines or from the sources. When changing the passaging method, it is necessary to first make sufficient stocks using the proposed method and then make changes to confirm that the characteristics after the change have not changed.