Rapid lysis to remove red blood cells

Rapid lysis to remove red blood cells

Equipment and reagents:
All supplies and reagents that come into direct contact with the cells must be sterile.
1. A test tube with a volumetric scale (conical bottom);
2. Pasteur pipettes (short and long);
3. Benchtop centrifuge;
4. Culture grade pure water;
5. 2 × Hanks balanced salt solution;
6. Storage medium containing 10% serum;

experimental method:
1. Centrifuge the primary cell suspension for 5 min at 1000 r/min and discard the supernatant;
2. Add 10 ml of culture-grade water to the cell pellet, and gently pipe back and forth several times with a pipette to resuspend the cells (Note: different cells have different anti-hypertension ability and control exposure time is less than 15 s);
3. Quickly add an equal volume of 2×Hanks balanced salt solution to restore tension, and gently mix the cell suspension with a Pasteur pipette;
4. Centrifuge again at 1000 r/min for 5 min to check for red blood cell pellets. Repeat this step if red blood cells are still present in large amounts. Otherwise resuspending the washed cells in storage medium;
5. After counting and detecting the viability of the cells, put them in an incubator for cultivation;

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