Category: Cell Culture
Protocol – Feeding Suspension Culture (Partial Replacement of Medium)
Replacement of culture medium from suspension culture is a bit tricky job as cells float in the medium. Ideally, cells are harvested from the medium by centrifugation and then resuspended in a fresh culture medium. However, in practice, a part of the culture medium is replaced with a fresh culture medium without harvesting the cells. This procedure is associated with the loss of cells.
Protocol – Thawing and Revival of Cryopreserved Cells
This protocol describes the revival process of cryopreserved adherent cells. The revival process involves rapid thawing of cryopreserved cells followed by removal of freezing medium which contains cryoprotectant, in this case, DMSO. The freezing medium can be removed immediately just after thawing the cells by centrifugation. Alternatively, cells are allowed to adhere to the culture dish followed by washing and addition of fresh growth medium.
Protocol – Passaging/Subculturing Suspension Culture
Suspension culture is passaged by diluting the existing culture. Since cells float in the medium in suspension culture, they are not treated with a trypsin-EDTA solution. To subculture a suspension cell line, a small amount of cell suspension from the existing culture is transferred to a culture dish containing fresh growth medium.