A 3-step method for preparing cryopreserved samples of apheresis products for post-thaw analysis yields a higher percentage of viable cells.
Transfusion
; 64(5): 866-870, 2024 May.
Article
em En
| MEDLINE
| ID: mdl-38606842
ABSTRACT
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 ANDMETHODS:
We developed a novel method whereby thawed samples were diluted step-wise to 12 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 110 dilution was required to obtain a desired cell concentration for flow cytometry testing resulting in a 120 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 120 yielded higher and more precise viability measurements compared to quickly washing samples to remove DMSO.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Remoção de Componentes Sanguíneos
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Criopreservação
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Sobrevivência Celular
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Citometria de Fluxo
Limite:
Humans
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article