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Cryopreservation of mobilized blood stem cells at a higher cell concentration without the use of a programmed freezer

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Abstract

Cryopreservation of peripheral blood stem cells (PBSC) mobilized by chemotherapy combined with or without granulocyte colony-stimulating factor (G-CSF) is an essential part of procedure for anti-cancer strategies. We evaluated whether a higher cell concentration (2×108/ml) without the use of a programmed freezer was acceptable for the storage of mobilized PBSC in an autologous setting. Mobilized PBSC were enriched to mononuclear cells (MNC) by Percoll separation and then frozen at cell concentrations of 2–5×107/ml (group I, n=20) or 2×108/ml (group II, n=44) without the use of a programmed freezer using 5% DMSO, 6% hydroxy ethyl starch, and 4% autologous serum or human albumin. CD34+ cells purified by ISOLEX300 were frozen at 2×107/ml (group III, n=22) using the same method. The median recovery rates of CD34+ cells and CFU-GM were, respectively, n.d. (not determined) and 88% in group I, 103 and 64% in group II, and 98 and 53% in group III. There was a statistical significance between the recovery rate of CFU-GM in group III and that in group I (p=0.02). The median percentage of cell viability after thawing in each group was 89, 87, and 75%, respectively. The median numbers of days after PBSCT to achieve a WBC of >1.0×109/l, an absolute neutrophil count of >0.5×109/l, and a platelet count of >50×109/l were, respectively, 11, 11 and 15 in group I; 12, 12 and 16 in group II; and 12, 12 and 27 in group III. These results suggest that enriched MNC from mobilized PBSC could be frozen at a higher cell concentration (2×108/ml) without the use of a programmed freezer, leading to reduction of the toxicities associated with infusion of thawed cells and of costly space required for cell storage.

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Acknowledgements

The authors are grateful to Ms. Yasuda for her excellent technical assistance and to the nursing staff of the pediatric ward at the University Hospital of Tokushima.

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Authors and Affiliations

  1. Department of Pediatrics, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, 890-8520, Kagoshima, Japan

    Yoshifumi Kawano

  2. Department of Pediatrics, University of Tokushima, Tokushima, Japan

    Yoshifumi Kawano, Tsutomu Watanabe, Takanori Abe, Hiroko Suzuya, Yasuhiro Okamoto, Atsushi Makimoto, Ryuji Nakagawa, Hiroyoshi Watanabe & Yoichi Takaue

  3. Department of Pediatrics, University of Malaya, Kuala Lumpur, Malaysia

    Chan Lee Lee

  4. Stem Cell Transplant Unit, National Cancer Center Hospital, Tokyo

    Yoichi Takaue

Authors
  1. Yoshifumi Kawano
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  2. Chan Lee Lee
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  3. Tsutomu Watanabe
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  4. Takanori Abe
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  5. Hiroko Suzuya
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  6. Yasuhiro Okamoto
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  7. Atsushi Makimoto
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  8. Ryuji Nakagawa
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  9. Hiroyoshi Watanabe
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  10. Yoichi Takaue
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Correspondence to Yoshifumi Kawano.

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Kawano, Y., Lee, C.L., Watanabe, T. et al. Cryopreservation of mobilized blood stem cells at a higher cell concentration without the use of a programmed freezer. Ann Hematol 83, 50–54 (2004). https://doi.org/10.1007/s00277-003-0817-8

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  • DOI: https://doi.org/10.1007/s00277-003-0817-8

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