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Frozen-thawed human blood monocytes respond reproducibly to activation stimuli: Implications for screening of BRMs

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Biotherapy

Abstract

The purpose of this study was to identify the optimal freezing conditions for human blood monocytes to allow their recovery and use forin vitro screening of activation stimuli. Human monocytes separated from buffy coats of healthy blood donors were suspended at a density of 1 × 107 cells/ml in freezing medium consisting of 70% medium: 20% fetal bovine serum: 10% DMSO frozen in a stepdown freezer, and stored at −180°C. Monocytes were thawed at different times up to 4 months later. Viability was >90%. Fresh monocytes from different donors and frozen monocytes thawed at different times were incubated with different concentrations of lipopolysaccharide, muramyl tripeptide, muramyl dipeptide, or lipopeptide. Tumoricidal activity and IL-1 production of fresh monocytes varied greatly among the 5 different preparations. In contrast, the frozen monocytes (thawed at different times) produced uniform levels of antitumor activity and IL-1 production. These results show that monocytes recovered from frozen storage maintain their ability to respond to activation stimuli in a uniform and reproducible manner. Thus, the use of frozen-thawed monocytes is recommended for screening of macrophage activating agents.

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Abbreviations

FBS:

fetal bovine serum

LPS:

lipopolysaccharide

IdUrd:

iododeoxyuridine

DMSO:

dimethyl sulfoxide

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Authors and Affiliations

  1. Department of Cell Biology, The University of Texas M.D. Anderson Cancer Center, 1515 Holcombe Boulevard, HMB 173, 77030, Houston, TX

    Teruhiro Utsugi, Dale Brown, Akihiko Nii & Isaiah J. Fidler

Authors
  1. Teruhiro Utsugi
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  2. Dale Brown
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  3. Akihiko Nii
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  4. Isaiah J. Fidler
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Utsugi, T., Brown, D., Nii, A. et al. Frozen-thawed human blood monocytes respond reproducibly to activation stimuli: Implications for screening of BRMs. Biotherapy 5, 301–308 (1992). https://doi.org/10.1007/BF02179048

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  • DOI: https://doi.org/10.1007/BF02179048

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