Human Biology

Human PBMC Isolation

  1. Dilute blood 1:1 with room temperature PBS.
  2. Carefully layer 2 volumes of diluted blood on top of 1 volume of lymphoprep in 50ml falcon tubes (ex: 25ml blood on top of 12.5ml lymphoprep).
    1. Make sure Pipet-Aid is on “medium up” and “slow down”
  3. Place tubes in aerosol containment buckets, and spin at 800xg (~1900rpm) for 20 minutes at room temp. Turn the brake OFF.
  4. Collect the PBMC layer at the interface between plasma and lymphoprep by slowly pipetting up in swirling motion with a glass Pasteur pipette on a pipet-aid.