X-Message-Number: 12362
Date: Thu, 2 Sep 1999 05:16:33 -0700 (PDT)
From: Doug Skrecky <>
Subject: long-term storage at -90 C

Authors
  Ayello J.  Semidei-Pomales M.  Preti R.  Hesdorffer C.  Reiss RF.
Institution
  Department of Pathology, Columbia-Presbyterian Medical Center, New York, NY
  10032, USA.
Title
  Effects of long-term storage at -90 degrees
  C of bone marrow and PBPC on cell recovery, viability, and clonogenic
  potential.
Source
  Journal of Hematotherapy.  7(4):385-90, 1998 Aug.
Abstract
  Autologous BM and PB HPC are usually stored from weeks to months until
  reinfusion after myeloablative chemotherapy. HPC have been stored for up to
  16 months at -90 degrees C, using a mixture of 5% DMSO, 6% hydroxyethyl
  starch (HES), and 4% HSA as a cryoprotectant. Long-term
  storage (LTS) has usually entailed rate-controlled freezing
  using 10% DMSO and preservation in liquid nitrogen. The effects of LTS at -90
  degrees C on the in vitro cell recovery, viability, and colony-forming
  unit-granulocyte macrophage (CFU-GM) clonogenic potential of autologous HPC
  that were not transplanted was studied. Sixteen BM and sixteen PB HPC had
  been cryopreserved for a median of 53 months (range 27-71) and 35 months
  (range 26-78), respectively. Samples of frozen HPC were thawed after 48 h,
  and the nucleated cell count, viability by trypan blue exclusion, and culture
  for CFU-GM were obtained. Following LTS, the cells were thawed and examined
  using the same assays. No difference in the median percentage recovery of
  nucleated cells was found in either the BM or PB HPC between the samples
  stored for 48 h and after LTS (5.73 x 10(9) versus 5.61 x 10(9) and 6.20 x
  10(9) versus 5.78 x 10(9), respectively). In addition, no difference in
  median percentage viability was found in either the BM or PB HPC sampled at
  48 h and at the end of LTS (75% versus 74% and 75% versus 76%, respectively).
  Finally, the median number of CFU-GM cultured from BM HPC at 48 h was 2.41 x
  10(5) (range 0.33-11.01 x 10(5)) and at the end of LTS was 1.93 x 10(5)
  (range 0.32-10.55), representing a median recovery of 93% (range 19%-308%).
  Similarly, the median number of CFU-GM cultured from PB HPC was 1.66 x 10(5)
  (range 0-50.57) and at the end of LTS was 0.93 x 10(5) (range 0-44.9),
  representing a median recovery of 80% (range 36%-165%). This difference in
  percentage recovery was not significant (p = 0.514). There was poor
  correlation between the number of nucleated cells harvested and the
  percentage recovery of nucleated cells, cell viability, or CFU-GM for either
  the BM or PB HPC. Similarly, there was poor correlation between the number of
  CFU-GM in the harvest and their percentage recovery following LTS for both BM
  and PB HPC. Finally, there was poor correlation between the
  storage time of the BM or PB HPC and the percentage recovery
  of nucleated cells, cell viability, and CFU-GM. These data suggest that LTS
  of HPC at -90 degrees C is not associated with decreased recovery of
  nucleated cells or in vitro viability and is associated with only a modest
  decrease in clonogenic potential. This indicates that
  storage of HPC at -90 degrees C for periods in excess of 3
  years is possible.

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