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A 3‐step method for preparing cryopreserved samples of apheresis products for post‐thaw analysis yields a higher percentage of viable cells

Strasburg, Dustin J. ; Sterner, Rosalie M. ; Va, Sildane ; Jacob, Eapen K. ; DiGuardo, Margaret A.

Transfusion (Philadelphia, Pa.), 2024-05, Vol.64 (5), p.866-870 [同儕審閱期刊]

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A 3‐step method for preparing cryopreserved samples of apheresis products for post‐thaw analysis yields a higher percentage of viable cells
著者: Strasburg, Dustin J. ; Sterner, Rosalie M. ; Va, Sildane ; Jacob, Eapen K. ; DiGuardo, Margaret A.
主題: Albumins ; Antigens, CD34 - analysis ; Apheresis ; Blood cells ; Blood Component Removal - methods ; Blood Preservation - methods ; CD34 ; CD34 antigen ; Cell death ; Cell Survival ; Cryopreservation ; Cryopreservation - methods ; Cryoprotectants ; Cryoprotective Agents - pharmacology ; Cryoprotectors ; Dextran ; Dextrans ; Dilution ; Enumeration ; Flow cytometry ; Flow Cytometry - methods ; Hematopoietic stem cells ; Hematopoietic Stem Cells - cytology ; Hemopoiesis ; HPC ; Humans ; Leukocytes ; Low molecular weights ; Lymphocytes ; Lymphocytes T ; Molecular weight ; post‐thaw ; Progenitor cells ; Sodium chloride ; viability ; Washing
所屬期刊: Transfusion (Philadelphia, Pa.), 2024-05, Vol.64 (5), p.866-870
描述: Background Standard flow cytometry protocols for CD34+ cell enumeration designed for fresh samples are not appropriate for cryopreserved products. Special protocols have been developed to remove the cryoprotectant by quickly washing a freshly thawed sample. Exposing cells to a large volume of hypotonic solution and subsequent washing process was hypothesized to cause lab‐induced cell death. Moreover, standard gating strategies must be altered to avoid reporting falsely high viabilities. Study Design and Methods We developed a novel method whereby thawed samples were diluted step‐wise to 1:2 by 3 additions of 1/3 sample volume using 1% Human Albumin in Dextran 40 (10% Low Molecular Weight Dextran in 0.9% NaCl) separated by 5 min between each addition. An additional 1:10 dilution was required to obtain a desired cell concentration for flow cytometry testing resulting in a 1:20 dilution. Results Twenty samples were tested simultaneously in a method comparison; the new method demonstrated significant increases in mean cell viabilities for white blood cells, hematopoietic progenitor cells, and T cells as well as reduced standard deviations for each parameter. Discussion Slow, step‐wise dilutions of freshly thawed samples of cryopreserved apheresis products to 1:20 yielded higher and more precise viability measurements compared to quickly washing samples to remove DMSO.
出版者: Hoboken, USA: John Wiley & Sons, Inc
語言: 英文
識別號: ISSN: 0041-1132
EISSN: 1537-2995
DOI: 10.1111/trf.17822
PMID: 38606842
資源來源: Wiley Online Library All Journals
Wiley Online Library Full Collection 2020

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A 3‐step method for preparing cryopreserved samples of apheresis products for post‐thaw analysis yields a higher percentage of viable cells

Strasburg, Dustin J. ; Sterner, Rosalie M. ; Va, Sildane ; Jacob, Eapen K. ; DiGuardo, Margaret A.

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