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Post-thaw yield and viability of Human Primary Immune cells

What can I do to improve the post-thaw yield and viability of cryopreserved Human Primary Immune cells?

We recommend the following thawing procedure:

1. Rapidly thaw the vial in a 37°C water bath. Remove the vial from the water bath as soon as

the contents are thawed, before all the ice has melted.

2. Decontaminate the vial by spraying with 70% ethanol and move it to a biosafety cabinet. All operations from this point onwards have to be carried out under strict aseptic conditions.

3. Carefully transfer the contents of the vial to a conical tube containing 5mL HBSS, Hank's Balanced Salt Solution without Ca2+ or Mg2+ (ATCC® 30-2213 ™) containing 10% Human AB serum or 10% Human Serum Albumin.

4. Rinse the vial with additional HBSS and transfer to the conical tube.

5. Bring the volume in the tube up to 10 mL with HBSS containing 10% HSA.

6. Take a sample for counting and viability assessment if needed.

7. Centrifuge the cell suspension at 300 x g for 5 minutes at room temperature.

8. Carefully aspirate the supernatant without disturbing the pellet, leave some supernatant behind.

9. Gently re‐suspend the cell pellet in an application-specific medium as needed. Generally RPMI‐1640 containing 10% human serum may also be used.

Date Created02/26/2018 08:02 AM
Date Updated02/26/2018 08:04 AM

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