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Suspension culture

A culture in which cells will multiply when suspended in growth medium. Suspension cultures are derived from cells that can survive and proliferate without attachment (anchorage independent); this ability is restricted to hematopoietic cells, transformed cell lines, and cells from malignant tumors. Cells that grow in suspension can be grown in any type of flask, plate, or Petri dish that, although sterile, need not be treated for cell attachment. Stirrer bottles are used when agitation is required to keep the cells in suspension. Agitation is usually by a suspended paddle containing a magnet, whose rotation is driven by a magnetic stirrer. The rotational speed must be kept low, 60 rpm, to avoid damage from shear stress. Generally, the pendulum design is preferable for minimizing shear. Suspension cultures can be set up as replicates or can be sampled repetitively from a side arm or the flask. They can also be used to maintain a steady-state culture by adding and removing medium continuously. A rapid stirring action for dissolving chemicals is available with any stirrer, but for the stirrer to be used for suspension culture (1) the motor should not heat the culture (use the rotating-field type of drive or a belt drive from an external motor); (2) the speed must be controlled down to 60 rpm; (3) the torque at low rpm should still be capable of stirring up to 10 L of fluid; (4) the device should be capable of maintaining several cultures simultaneously; (5) each stirrer position should be individually controlled; and (6) a readout of rpm should appear for each position. It is preferable to have a dedicated stirrer for culture work The viscosity of a culture medium is influenced mainly by the serum content and in most cases will have little effect on cell growth. Viscosity becomes important, however, whenever a cell suspension is agitated (Higher viscosity higher damage to cells). Foaming is another problem in suspension culture arise due to agitation of culture for this purpose antifoaming agent used. Calcium is reduced in suspension cultures in order to minimize cell aggregation and attachment The choice of BSS also is dependent on whether the solution will be used for suspension culture of adherent cells. S-MEM, based on Eagles Spinner salt solution, is a variant of Eagles [1959] minimum essential medium that is deficient in Ca2+ in order to reduce cell aggregation and attachment. Suspension cultures have a number of advantages for example, trypsin treatment is not required so subculture is quicker and less traumatic for the cells, and scale-up is easier. Replacement of the medium (feeding) is not

usually carried out with suspension cultures, and instead, the culture is either diluted and expanded, diluted and the excess discarded, or the bulk of the cell suspension is withdrawn and the residue is diluted back to an appropriate seeding concentration.