The invention discloses a method for separating
peripheral blood mononuclear cells from a repayment-
free blood donation
whole blood collection device, which comprises the following steps: clamping two side tube walls by using a tube clamp under a sterile condition,
cutting open upper and lower end tube orifices of the two side tube walls by using sterile scissors, and reserving a section of long tube on each of the two side tube walls; using an
injector to forward flush the white filter and backward flush the white filter with the flushing fluid, collecting the liquid obtained by backward flushing, centrifuging, sucking and discarding the supernatant after centrifuging, re-suspending cells with normal
saline, adding into another two centrifugal tubes filled with the same amount of
Ficoll, and continuing centrifuging; and after
centrifugation is finished, sucking and abandoning supernatant, sucking the albuginea layer above the
Ficoll into a new
centrifuge tube, supplementing normal
saline, sucking and abandoning supernatant after
centrifugation, and manually counting by adopting a
blood cell counting plate after the normal
saline is resuspended. According to the method disclosed by the invention, the mononuclear cells can be aseptically extracted from the white filter of a blood donation and collection pipeline and cultured
in vitro, and the cells cultured
in vitro are used as DC-CIK treatment cells.