Evaluation of Collection and Processing Conditions for Gene Expression Analysis Using Human Myeloid Cells.
Biopreserv Biobank
; 2024 Mar 22.
Article
in En
| MEDLINE
| ID: mdl-38526566
ABSTRACT
Background:
The population of blast cells among peripheral blood mononuclear cells (PBMCs) obtained from patients is a desirable specimen for analyzing gene expression in diseases including acute myeloid leukemia. Although the enrichment of blast cells often needs to be performed at a central laboratory, acceptable conditions for sample transport from clinical sites remain to be established.Methods:
We evaluated storage temperature, duration, and tube type before initiating sample processing for the analysis of cluster of differentiation (CD)33+ myeloid cells among PBMCs as an alternative to CD34+/CD33+ blast cells.Results:
CD33+ myeloid cells were successfully purified by MACS. The cell viability and the RNA integrity were sustained during storage up to 48 hours before sample processing. Storage at 4°C had minimal effects on gene expression, whereas storage at room temperature induced the senescence pathway, characterized by the expression of stress-inducible genes. A CPT tube was also better than an ethylenediaminetetraacetic acid tube for minimizing gene expression change.Conclusions:
Our study provided important clues for establishing a sample handling approach for gene expression analysis with purified cell fractions from human PBMCs. To keep the variation of gene expression to a minimum, samples should be delivered at 4°C within 48 hours before processing.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Language:
En
Journal:
Biopreserv Biobank
/
Biopreserv. biobank. (Online)
/
Biopreservation and biobanking (Online)
Year:
2024
Document type:
Article
Affiliation country:
Japón
Country of publication:
Estados Unidos