How can I isolate human platelets by depletion of unwanted cells?

Below you find a customer protocol for the isolation of platelets by depletion of unwanted cells:

  1. Start with 9.0 mL of citrate-stabilized blood.
  2. Centrifuge for 10 min at 200xg at room temperature. This yields a thrombocyte-rich supernatant with 3x107 erythrocytes, 1.4x106 lymphocytes, and 5x108 platelets.
  3. Combine 350 µL of this supernatant with 100 µL of CD45 MicroBeads, human and 100 µL of CD235a (Glycophorin A) MicroBeads, human.
  4. Incubate at room temperature (25 °C) for 6 min. In the mean time, prepare a MS Column and MiniMACS Separator or OctoMACS Separator.
  5. Apply the sample to the MS Column.
  6. Wash three times with 500 µL PBS containing 2 mM citrate and 0.5% BSA. The flow-through will contain the untouched platelets. If a higher concentration of platelets is required, use only one washing step.