Method for purifying cord blood mononuclear cells
A technology of nuclear cells and blood cells, applied in the field of biomedicine, can solve the problem of low purity of umbilical cord blood mononuclear cells, and achieve the effects of large number, high purity and high viability.
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Embodiment 1
[0056] Divide 60ml of umbilical cord blood samples into 50ml centrifuge tubes, centrifuge at 500g for 10min to obtain blood cells and plasma, and collect the plasma for later use.
[0057] Preheat hydroxyethyl starch in a water bath at 30°C for 15 minutes, then add the preheated hydroxyethyl starch to blood cells for precipitation, the volume ratio of blood cell sedimentation to hydroxyethyl starch is 3:1, mix well and let stand 3min, then add normal saline, the volume ratio of normal saline to blood cell sediment is 5:3, then mix and let stand for 30min.
[0058] The cell suspension in the upper layer was taken and centrifuged at a centrifugal force of 500 g for 8 min to obtain lymphocyte precipitates.
[0059] Mix the lymphocyte pellet with the spare plasma to obtain a lymphocyte suspension, and then slowly add it to the Ficoll liquid surface to make a clear interface. The volume ratio of the lymphocyte suspension to the Ficoll liquid surface is 1:1, and then use 400g centri...
Embodiment 2
[0063] Divide 60ml of umbilical cord blood samples into 50ml centrifuge tubes, centrifuge at 500g for 10min to obtain blood cells and plasma, and collect the plasma for later use.
[0064] Preheat hydroxyethyl starch in a water bath at 30°C for 15 minutes, then add the preheated hydroxyethyl starch to blood cells for precipitation, the volume ratio of blood cell sedimentation to hydroxyethyl starch is 3:1, mix well and let stand 3min, then add normal saline, the volume ratio of normal saline to blood cell sediment is 5:3, then mix and let stand for 30min.
[0065] The cell suspension in the upper layer was taken and centrifuged at a centrifugal force of 500 g for 8 min to obtain lymphocyte precipitates.
[0066] Mix the lymphocyte pellet with the spare plasma to obtain a lymphocyte suspension, and then slowly add it to the Ficoll liquid surface to make a clear interface. The volume ratio of the lymphocyte suspension to the Ficoll liquid surface is 1:1, and then use 400g centri...
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