Methods and Materials: Trituration

(Cell dispersion through mild pumping action)

This can be a crucial procedure. It serves to break up the tissue fragments following incubation in the dissociation mix. If done too vigorously, cells will be destroyed lowering viability; too weakly and tissue fragments will be left intact lowering yield. Gentle trituration, using a 10 ml pipette, constitutes filling and emptying the barrel at a rate of about 3.0 ml per sec. You can best determine a suitable rate for your tissue through trial and error. Avoid bubbling the cell suspension.