Thawing Recommendations

NOTE: This protocol is a general guideline only, always follow the protocol at your institution

Liquid Nitrogen Storage: If the Cryostore freezing bag has been stored in the liquid phase of the liquid nitrogen (LN) tank and is to be removed for thawing, move the container into the vapor phase of the tank for at least 4 hours before removing from the storage tank. This will allow LN that may have penetrated the bag to gassify and escape more slowly, helping reduce the chance of bursting on rewarming.

Rewarming: We recommend leaving the bag in the cassette until just before immersion in a water bath. If the cassette cannot be easily removed form the bag, warm the cassette to allow the frost holding the cassette closed to melt.

The water bath should be cleaned, disinfected, and heated to 38ºC±2º. Sterile water is recommended.

During warming, observe the bag for any problems, such as breakage, leakage or swelling. Watch the bag carefully for expansion, indicating LN infiltration. If this occurs, puncture the spike port to release pressure and prevent bursting. Using an aseptic technique may help save the sample.

The bag is considered thawed when no ice crystals remain in the bag. It should be removed from the ice bath immediately, as some cryoprotectants may alter cells at elevated temperatures.

The Cryostore bag is provided with hermetically sealed spike ports. To open these, hold the bag at the top seal and twist off the top portion of the spike port, breaking the seal at the red arrow. The top portion will tear off at the red arrow, exposing the sterile port. Puncture with any standard blood spike.

OriGen Biomedical Europe
Drottninggatan 13
25221 Helsingborg
Sweden
Tel: +46 46 772 433
Fax: +46 42 146 074

Key list: Blood banking, cord blood storage, cryopreservation,stem cell storage, cord blood freezing, blood component freezing, hematopoetic