Freezing Information

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

Sealing:

To seal the donor tube, a dielectric or RF sealer such as the SEBRA model 2600/1105 Hand Held sealer is recommended, Click here for a link to the SEBRA website. The sealer should be operated and maintained according to manufacturer's instructions to ensure optimal performance.

Seal the donor tube so that the trimmed seal does not project above the spike port tops (yellow arrow). Keep the sealer jaws perpendicular to the tube being sealed, and make the seal parallel to the face of the bag. After sealing, press the bag gently with the flat of your hand and inspect the seal for leaks. Do NOT seal tube in the same place twice. If the seal has a leak, seal again below it.

When the seal is acceptable, cut off the donor tube just above the seal. Be sure that the donor tube stub does not extend past the tops of the spike port caps, as shown to the right.

Sample segments may be created by placing additional seals in the filled donor tube at suitable intervals. To provide the maximum degree of sample traceability, leave the sample links attached to the bag during storage. If sample links are not needed, the donor tube may be cut above the upper seal using scissors.

NOTE: Carefully check for air in the seal area as freezing may cause increased pressure in the seal. Check the seals for leakage by gently applying pressure on the container.

Freezing Protocol:

Controlled rate freezing to below -40ºC is highly recommended to help prevent breaking of the bag in storage. The use of a fixed rate freezing protocol is strongly suggested to freeze the bags before placing into the vapor phase of a liquid nitrogen storage tank.

1. Prepare: Dry the freezing bag and the interior of a freezing cassette. It is recommended that only one bag be stored per cassette.

2. Freeze: Freeze the bag in the cassette using a controlled rate freezing protocol. Freezing the bag to below -40ºC.

DMSO Cryoprotectant Procedure* Click here for link

DMSO/Pentastarch Cryoprotectant Procedure* Click here for link

*The above procedures were presented at the ISCT 2001 meeting; these sample procedures are for reference only.

Following completion of the freezing program, remove the cassettes from the freezing chamber and place them into the storage racks within the vapor phase of the nitrogen storage tank. Transfer the bags to the storage tank as quickly as possible.

A uniform thickness of the product is recommended for best freezing results. The fill volume is dependent on the way that the bag is to be stored in liquid nitrogen. This slight variance is best determined by the user.

Cryostore Freezing Bags are fragile when frozen, please handle carefully.

While the Cryostore Freezing bags are suitable for direct immersion in the liquid phase of the nitrogen tank, it is recommended that the container be stored in the vapor phase of the tank to avoid possible cross contamination.

Never move room temperature products into Liquid Nitrogen, as bursting of the bag is very probable. Do not immerse the CryoStore freezing bag directly into liquid nitrogen from the temp control freezer. Allow to cool to vapor phase temperatures first. Anything that causes extreme thermal shock can result in damage to the Cryostore bag.

Always follow your institutional protocol and all safety procedures when working with liquid nitrogen.

Cassettes

The Cryostore bag has been designed to fit into existing, standard cassettes. Check to be sure that the selected bag fits comfortably in the selected cassette. We recommend using one bag per cassette.

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