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Post-thaw yield and viability of ATCC® PCS-800-012 ™ Human Primary Bone Marrow CD34+ cells

What can I do to improve the post-thaw yield and viability of cryopreserved ATCC® PCS-800-012 ™, Human Primary Bone Marrow CD34+ cells?

We recommend the following thawing procedure:

  • Rapidly thaw the cryovial in a 37°C water bath (1-2 min). Remove the vial before all the ice is thawed.
  • Decontaminate the cryovial with 70% ethanol and move it to a biosafety cabinet and keep on ice.
  • While on ice, transfer contents of the vial to a 15mL conical tube.
  • Rinse the cryovial with 1mL of ice-cold thawing media (Hank's Balanced Salt Solution without Ca2+ or Mg2+ (ATCC® 30-2213 ™), supplemented with 10% Fetal Bovine Serum (ATCC® 30-2020 ™) and slowly transfer to the cells to the conical tube.
  • Slowly pipet 10X the vial volume of thawing media into the conical tube.
  • Take a sample for counting and viability assessment.
  • Centrifuge the cells at 4°C at 300 x g for 5 minutes to pellet cells.
  • Aspirate the supernatant carefully, leaving a few mLs behind.
  • Re-suspend cell pellet in the assay media of your choice.

While some reports (Ref) suggest increased viability using warm thawing media, ATCC has routinely achieved viability of over 80% as determined by trypan blue exclusion assays immediately post-thaw, using this protocol.

Date Created08/27/2014 07:14 AM
Date Updated01/12/2017 09:27 AM

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