Purification of human peripheral blood mononuclear cells by density retardation

Reagents and equipment:
1. Anticoagulant: 100mmol / L ethylenediaminetetraacetic acid or 38g / L sodium citrate;
2. Hepes buffered saline (HBS): 8.5 g / L NaCl, 10 mmol / L Hepes-NaOH, pH 7.4;
3. Lymphocyte separation solution (such as LymphoprepTM or NycoPrepTM1.077) or OptiPrepTM, diluted with Hepes buffered saline (5 volumes and 17 volumes, respectively);
4. Benchtop low speed centrifuge;
5. Pasteur pipettes (short and long);
6. 5-10ml size syringe and metal trocar;

experimental method:
1. Collect venous blood with ethylenediaminetetraacetic acid (2mmol/L final concentration) or sodium citrate (38g/L final concentration) as an anticoagulant;
2. Dilute the blood with an equal volume of Hepes buffered saline and gently invert it several times;
3. Using a plastic Pasteur pipette, add 6 ml of diluted blood to the selected medium of 3 ml of the centrifuge tube;
4. Centrifuge at 700g for 15min (LymphoprepTM) or 700g for 20min (NycoPrepTM 1.077 or iodixanol solution) at 20 ° C;
5. Slow down the rotor with a slow deceleration program or without braking;
6. Collect cells at the interface using a Pasteur pipette;
7. Dilute with 2 volumes of Hepes buffered saline and centrifuge at 200g-500g for 15 min to collect the cells;

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  Purple onions and yellow onions also have a big difference in taste. Purple onions have a lighter spicy taste and a slightly sweet taste. They can be eaten raw, but the yellow onions are particularly spicy. They cannot be eaten raw and are suitable for frying. Eat after making or making soup. In addition, the purple onion has relatively thick flesh and high juice content, while the yellow onion has very thin flesh quality and low juice content.

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