Is it possible to store red cells from "problem patients" for prolonged periods without using liquid nitrogen?

The following question was submitted to the e-network for discussion: Is it possible to store red cells from "problem patients" for weeks to months (e.g. sensitized cells, or red cells in general), without using liquid nitrogen? Does anyone have experience with storing liquid samples for extended time periods, which would allow for building a collection of samples (with sera, of course) to test new techniques (e.g. column methods)?

To which the following replies were received.

1. In regard to liquid storage of whole blood, in an Editorial on "Detecting alloantibodies in patients with autoantibodies" that appeared during January of 1999 in Transfusion (volume 39 pages 6-10) it was stated that RBCs can be stored in the liquid state for about 2 months. This is what has been done at UCLA in the past and apparently the cells maintain their antigen integrity over this time period.
   
   
2. In addition, at the LAC+USC Medical Center, extended storage of red cell samples has been accomplished by placing the red cells in an Alsever's solution. It can be purchased from Immucor /Gamma.

ADDENDA Jan. 3, 2001

3. The only liquid cells that we store for any extended period are red cells that we utilize for special panels, such as, cord panels, Bg panels, etc. We remove the plasma or serum and then make a 3% cell suspensions in Alsever's and store these at 4C for 4 weeks (but no longer than 66 days from the date the sample was drawn). We do store frozen cell samples and the procedure is as follows:
   1. For each volume of red cells(washed 1x with saline, add 2 volumes of glycerol(drop wise), mix by inversion and freeze.
   2. Thaw cells at RT and mix by inversion.
   3. For each volume of red cells, add equal volume of 9.0% NaCl, mix and equilibrate at RT for one minute.
   4. Fill tube with 2.5% NaCl, mix and invert.
   5. Centrifuge and remove supernatant.
   6. Fill tube again with 2.5% NaCl and mix and invert.
   7. Centrifuge and remove supernatent
   8. Wash 2x with 0.9% saline or until there is no hemoglobin in the supernatent.

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