The invention provides a
separation method for separating
peripheral blood mononuclear cells, and particularly relates to the field of biological medicines. The
separation method comprises the following steps: S1, preparation of a separation tube: firstly, adding 2-6 ml of
Percoll or polysucrose or
meglumine diatrizoate
cell separation liquid with the density of 1.075-1.0796 g / ml into a centrifugal tube, sucking 0.5-1.5 ml of separation gel with the density of 1.06-1.07 g / ml, adding the separation gel into a tube opening of the centrifugal tube, and carrying out horizontal centrifuging for 1-3minutes at
room temperature under the
centrifugal force of 800-1200 g, so that the separation gel forms an
isolation layer on the liquid surface of the
Percoll or polysucrose or
meglumine diatrizoatecell separation liquid, and preparation of the separation tube is finished; and S2, separation of the
peripheral blood mononuclear cells: adding 2-6 ml of
anticoagulant whole blood into the preparedseparation tube, and carrying out horizontal centrifuging for 8-12 minutes at
room temperature under the
centrifugal force of 800-1200 g; sucking and discarding the uppermost liquid to remove
cell fragments and platelets, directly pouring the liquid above the
isolation layer into a collection tube, carrying out centrifuging for 4-6 minutes at
room temperature under the
centrifugal force of 600-1000 g, and resuspending the cells by using PBS to obtain the
peripheral blood mononuclear cells. According to the
separation method, it can be ensured that the
anticoagulant whole blood can be absolutely not mixed with a
cell separation medium after being added, and other cells are not polluted when the
peripheral blood mononuclear cells are harvested.