Recommendations for Thawing Cell Line Cryo-Pellets

If you prefer receiving the cell line as a pellet frozen in a cryo-protectant solution, please note that we recommend handling the cryo-vial immediately upon arrival in your laboratory in the following manner

  • Thawing and culturing the cells in the appropriate culture medium immediately
  • If not convenient, store immediately at -80° or better yet,
  • Store immediately for 8 to 24 hours at -80° and then transfer to liquid nitrogen storage

READ PRODUCT DESCRIPTION INSERT PAGE THAT WAS ENCLOSED WITH THE CRYO-VIAL FOR RECOMMENDED CULTURE CONDITIONS. AN ELECTRONIC PDF VERSION OF THE INSERT IS ALSO AVAILABLE AT OUR WEB SITE

THAWING PROTOCOL

  1. Warm the cryo-vial to room temperature in warm-block or water bath (25° ~ 30° C) taking care not to allow moisture to reach the vial closure risking contamination of the contents.
  2. As soon as the content of the vial is liquefied, transfer the cells with sterile technique into 40 ml of culture medium complete with serum as recommended. The purpose of this step is to dilute the cryo-protectant to a safe, non-toxic concentration for the cells.
  3. Centrifuge at 500 rpm for 5 minutes at room temperature to pellet the cells. Carefully remove supernatant above cells with a sterile pipette.
  4. Re-suspend cells in 10 ml culture medium complete with serum as recommended on the Product Description page.
  5. Plate 2 ml of cells on a sterile cell culture plate. Check density and adjust plating volumes to achieve 70% to 80% confluence in the culture vessels of your choosing.

Questions regarding these policies or technical cell culture queries related to the product may be referred to John Aletta, Director of Research (jmaletta@ch3biosystems.com).

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