ainer. Keep the canister in the vapor phase, just above the surface of the LN2 for 5-10 minutes before proceeding.
Note: If two different HEMACORD products are stored in the LN2 container at the same time, open one canister at a time with the canister opening tool as described above. Carefully check the ID number on the labels attached to the canister and the product, respectively. Close the canister and leave it in the vapor phase for 5-10 min. before proceeding.
a.Open canister with the canister opening tool as described above.
b.Work carefully to avoid damaging the frozen plastic product bag. Remember that plastic at this temperature is very brittle and breaks easily.
c.Examine the bag for breaks or cracks and document this inspection on the appropriate form.
d.Remove the HEMACORD from the canister.
Caution! Do not handle the plastic bags at liquid nitrogen temperature with the tongs intended for metal canisters, as this may rip the bag. Do not allow the product or tubing to bend as it may crack.
e.Put the HEMACORD inside a zipper-locked plastic bag, let the air out and close the bag. Place the bag with the HEMACORD in a warm water bath at approximately 38°C.
f.To accelerate and homogenize thawing, carefully agitate the product bag in the water and gently knead its contents.
g.Inspect and watch for leakage. If product leaks out into the zipper-locked bag, find the site of the leak in the freezing bag and position the bag so as to prevent further escape of product. While maintaining the bag in that position, finish thawing the product. (See Section 5 for emergency product recovery in the event of a container failure.)
h.As soon as the bag’s contents become slushy, remove the bag from the water bath and place it inside a biological safety cabinet.
3.3 Connecting the Freezing Bag to the Transplant Set
The procedure to restore the osmolarity of the HPC-C cell suspension, and either remove the supernatant with DMSO or simply dilute the thawed HEMACORD, is assisted by a sterile, empty, transplant bag set designed with two spike tubes to drain both compartments of the freezing bag (see Figure 4: “Cell Wash/Infusion Bag Set”). The Cell Wash/Infusion Bag Set is included with this shipment.
Note: The following procedure must be done in a biological safety cabinet.
Figure 4. Cell Wash/Infusion Bag Set
a.Close all clamps on the Cell Wash/Infusion Bag Set.
b.Remove the HEMACORD freezing bag from the zipper-locked bag.
c.Disinfect the covers of both ports of the freezing bag with iodine.
d.Using a clean and disinfected scissors, cut off the hermetically sealed covers of the freezing bag’s spike ports (Figure 5).
Figure 5.
e.Disinfect the cut surfaces of the spike port area of the freezing bag using iodine swab sticks (Figure 6).
Figure 6.
f.Insert the spikes of the Cell Wash/Infusion Bag Set into the ports of the freezing bag.
g.Label the transplant bag (shown in Figure 4) with HEMACORD ID number and the name of the recipient (or label according to local standard operating procedure).
3.4 Reconstitute (dilute) the thawed HEMACORD
The amount of thawing solution used for HEMACORD is at least 5 times the volume of the frozen product including the cryoprotectant. For example, 25 mL products are diluted to 170 mL total, and thus, a volume of 145 mL of thawing solution is required to make the final volume of 170 mL in a transplant bag.
a.Add first a volume of thawing solution equal to the volume of thawed HEMACORD (1:1 r |
