PBMC separation method
A separation liquid, RPMI1640 technology, applied in the field of separation of PBMC, can solve the problems of inconspicuous stratification, large amount of Ficoll separation liquid, and inability to recover plasma, etc., and achieve the effect of shortening separation time, improving yield and purity, and saving time and cost
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[0023] Example 1
[0024] Transfer 20 mL of healthy human anticoagulated peripheral blood to a 50 mL centrifuge tube, centrifuge at 18°C and 1000 g for 10 min; the peripheral blood in the centrifuge tube is divided into two layers, the upper layer is the first plasma layer, and the lower layer is the blood cell layer. The first plasma layer was centrifuged at 18° C. and 2500 g for 10 min, and the supernatant was filtered through a 0.22 μm filter to remove red blood cells, and the filtrate was plasma. Plasma is rich in a large number of protein factors, which can be used for subsequent cell culture experiments.
[0025] Dilute the blood cell layer and normal saline at a volume ratio of 1:1.5, and mix thoroughly to form a blood cell dilution solution. The blood cell diluent was added to the top of the Ficoll separation solution in the centrifuge tube, wherein the volume ratio of the Ficoll separation solution to the blood cell dilution solution was 1:2, and centrifuged for 30...
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[0027] Example 2
[0028] Transfer 20mL of healthy human anticoagulated peripheral blood to a 50mL centrifuge tube, centrifuge at 18°C and 800g for 10min; the peripheral blood in the centrifuge tube is divided into two layers, the upper layer is the first plasma layer, and the lower layer is the blood cell layer. The first plasma layer was centrifuged at 18° C. and 3000 g for 10 min, and the supernatant was filtered through a 0.22 μm filter to remove red blood cells, and the filtrate was plasma. Plasma is rich in a large number of protein factors, which can be used for subsequent cell culture experiments.
[0029] The blood cell layer and normal saline were diluted 1:2 times by volume, and mixed well to form a blood cell dilution solution. The blood cell dilution solution was added above the Ficoll separation solution in the centrifuge tube, wherein the volume ratio of the Ficoll separation solution to the blood cell dilution solution was 1:2, and centrifuged for 25 min at ...
Example Embodiment
[0031] Example 3
[0032] Transfer 20 mL of healthy human anticoagulated peripheral blood to a 50 mL centrifuge tube, centrifuge at 18°C and 600 g for 10 min; the peripheral blood in the centrifuge tube is divided into two layers, the upper layer is the first plasma layer, and the lower layer is the blood cell layer. The first plasma layer was centrifuged at 18° C. and 3000 g for 10 min, and the supernatant was filtered through a 0.22 μm filter to remove red blood cells, and the filtrate was plasma. Plasma is rich in a large number of protein factors, which can be used for subsequent cell culture experiments.
[0033] Dilute the blood cell layer and normal saline at a volume ratio of 1:2.5, and mix thoroughly to form a blood cell dilution solution. The blood cell diluent was added to the top of the Ficoll separation solution in the centrifuge tube, wherein the volume ratio of the Ficoll separation solution to the blood cell dilution solution was 1:2, and centrifuged for 20 ...
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