Aberrant determination of phenotypic markers in chronic lymphocytic leukemia (CLL) lymphocytes after cryopreservation

•Cryopreservation is a common laboratory technique used for long-term storage of blood cells.•Cryopreservation alters the detection of epitopes by flow cytometry.•Care must be taken when interpreting expression of epitopes on thawed samples. The cryopreservation of peripheral blood mononuclear cells...

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Bibliographic Details
Published in:Experimental hematology 2018-07, Vol.63, p.28-32.e1
Main Authors: Thurgood, Lauren A., Lower, Karen M., Macardle, Cindy, Kuss, Bryone J.
Format: Article
Language:English
Subjects:
Antigens, CD - analysis
Antigens, Neoplasm - analysis
Biomarkers, Tumor - analysis
Blood Preservation - methods
Cryopreservation
Epitopes - analysis
False Negative Reactions
Flow Cytometry - methods
Humans
Immunophenotyping - methods
Leukemia, Lymphocytic, Chronic, B-Cell - blood
Leukemia, Lymphocytic, Chronic, B-Cell - immunology
Leukemia, Lymphocytic, Chronic, B-Cell - pathology
Receptors, CXCR - analysis
Time Factors
Tumor Cells, Cultured
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Summary:•Cryopreservation is a common laboratory technique used for long-term storage of blood cells.•Cryopreservation alters the detection of epitopes by flow cytometry.•Care must be taken when interpreting expression of epitopes on thawed samples. The cryopreservation of peripheral blood mononuclear cells (PBMCs) is a routine research laboratory process, enabling long-term storage of primary patient blood samples. Retrospective analysis of these samples has the potential to identify markers that may be associated with prognosis and response to treatment. To draw valid biological conclusions from this type of analysis, it is essential to ensure that any observed changes are directly related to the pathology of the disease rather than the preservation process itself. Therefore, we have investigated 15 cell surface markers that are relevant to chronic lymphocytic leukemia (CLL) on matched fresh and thawed samples to determine the effect of cryopreservation on their detection. We found that the number of CLL cells positive for the markers CD22, CD40, CD49d, CD54, CD69, and CXCR3 was decreased significantly after cryopreservation. In addition, the mean fluorescence intensity of 10 of the 15 markers changed significantly after cryopreservation. These findings demonstrate that care must be taken when interpreting this type of analysis on thawed samples.
ISSN:0301-472X
1873-2399
DOI:10.1016/j.exphem.2018.04.003