The Cytokinesis-Block Micronucleus Assay on Human Isolated Fresh and Cryopreserved Peripheral Blood Mononuclear Cells

The cytokinesis-block micronucleus (CBMN) assay is a standardized method used for genotoxicity studies. Conventional whole blood cultures (WBC) are often used for this assay, although the assay can also be performed on isolated peripheral blood mononuclear cell (PBMC) cultures. However, the standardization of a protocol for the PBMC CBMN assay has not been investigated extensively. The aim of this study was to optimize a reliable CBMN assay protocol for fresh and cryopreserved peripheral blood mononuclear cells (PBMCS), and to compare micronuclei (MNi) results between WBC and PBMC cultures. The G0 CBMN assay was performed on whole blood, freshly isolated, and cryopreserved PBMCS from healthy human blood samples and five radiosensitive patient samples. Cells were exposed to 220 kV X-ray in vitro doses ranging from 0.5 to 2 Gy. The optimized PBMC CBMN assay showed adequate repeatability and small inter-individual variability. MNi values were significantly higher for WBC than for fresh PBMCS. Additionally, cryopreservation of PBMCS resulted in a significant increase of MNi values, while different cryopreservation times had no significant impact. In conclusion, our standardized CBMN assay on fresh and cryopreserved PBMCS can be used for genotoxicity studies, biological dosimetry, and radiosensitivity assessment.

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Whole blood can be used as an alternative to isolated peripheral blood mononuclear cells to measure in vitro specific T-cell responses in human samples

Journal of Immunological Methods ◽

10.1016/j.jim.2020.112940 ◽

2021 ◽

pp. 112940

Author(s):

Philippe Moris ◽

Aurélie Bellanger ◽

Opokua Ofori-Anyinam ◽

Erik Jongert ◽

Juan-Pablo Yarzabal Rodriguez ◽

...

Keyword(s):

T Cell ◽

Peripheral Blood Mononuclear Cells ◽

Whole Blood ◽

Peripheral Blood ◽

Mononuclear Cells ◽

T Cell Responses ◽

Peripheral Blood Mononuclear ◽

Cell Responses ◽

Blood Mononuclear Cells

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Familial differences in endotoxin-induced TNF release in whole blood and peripheral blood mononuclear cells in vitro; relationship to TNF gene polymorphism

Journal of Endotoxin Research ◽

10.1177/096805199500200103 ◽

1995 ◽

Vol 2 (1) ◽

pp. 19-25 ◽

Cited By ~ 17

Author(s):

H.H.F. Derkx ◽

K.F. Bruin ◽

C.V. Jongeneel ◽

L.P. de Waal ◽

B.M.N. Brinkman ◽

...

Keyword(s):

Gene Polymorphism ◽

Peripheral Blood Mononuclear Cells ◽

Whole Blood ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells ◽

Tnf Gene ◽

Familial Differences

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Comparative in vitro stimulation with lipopolysaccharide to study TNFα gene expression in fresh whole blood, fresh and frozen peripheral blood mononuclear cells

Journal of Immunological Methods ◽

10.1016/j.jim.2010.03.006 ◽

2010 ◽

Vol 357 (1-2) ◽

pp. 33-37 ◽

Cited By ~ 19

Author(s):

Jiwang Chen ◽

Anke H. Bruns ◽

Helen K. Donnelly ◽

Richard G. Wunderink

Keyword(s):

Gene Expression ◽

Peripheral Blood Mononuclear Cells ◽

Whole Blood ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells

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The effects of Malassezia on pro-inflammatory cytokine production by human peripheral blood mononuclear cells in vitro

Medical Mycology ◽

10.1046/j.1365-280x.1998.00137.x ◽

1998 ◽

Vol 36 (2) ◽

pp. 97-106 ◽

Cited By ~ 6

Author(s):

S. KESAVAN ◽

C. E. WALTERS ◽

K. T. HOLLAND ◽

E. INGHAM

Keyword(s):

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Inflammatory Cytokine ◽

Mononuclear Cells ◽

Cytokine Production ◽

Human Peripheral Blood ◽

Peripheral Blood Mononuclear ◽

Inflammatory Cytokine Production ◽

Blood Mononuclear Cells

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Measurement of ERK1/2 isoform distribution in peripheral blood mononuclear cells derived from whole blood from patients with different neuropsychiatric disorders

Pharmacopsychiatry ◽

10.1055/s-0035-1558049 ◽

2015 ◽

Vol 48 (06) ◽

Author(s):

I Kraus ◽

D Besong Agbo ◽

M Otto ◽

H Klafki ◽

J Wiltfang

Keyword(s):

Peripheral Blood Mononuclear Cells ◽

Whole Blood ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Neuropsychiatric Disorders ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells

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CD8+ T cells from feline immunodeficiency virus (FIV) infected cats suppress exogenous FIV replication of their peripheral blood mononuclear cells in vitro

Archives of Virology ◽

10.1007/s00705-002-0827-1 ◽

2002 ◽

Vol 147 (8) ◽

pp. 1517-1529 ◽

Cited By ~ 14

Author(s):

T. Hohdatsu ◽

T. Sasagawa ◽

A. Yamazaki ◽

K. Motokawa ◽

H. Kusuhara ◽

...

Keyword(s):

T Cells ◽

Peripheral Blood Mononuclear Cells ◽

Cd8 T Cells ◽

Feline Immunodeficiency Virus ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Peripheral Blood Mononuclear ◽

Immunodeficiency Virus ◽

Blood Mononuclear Cells

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Potential Role and Impact of Peripheral Blood Mononuclear Cells in Radiographic Axial Spondyloarthritis-Associated Endothelial Dysfunction

Diagnostics ◽

10.3390/diagnostics11061037 ◽

2021 ◽

Vol 11 (6) ◽

pp. 1037

Author(s):

Patricia Ruiz-Limon ◽

Maria L. Ladehesa-Pineda ◽

Clementina Lopez-Medina ◽

Chary Lopez-Pedrera ◽

Maria C. Abalos-Aguilera ◽

...

Keyword(s):

Endothelial Dysfunction ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Axial Spondyloarthritis ◽

Endothelial Injury ◽

In Vitro Studies ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells

Endothelial dysfunction (ED) is well known as a process that can lead to atherosclerosis and is frequently presented in radiographic axial spondyloarthritis (r-axSpA) patients. Here, we investigated cellular and molecular mechanisms underlying r-axSpA-related ED, and analyzed the potential effect of peripheral blood mononuclear cells (PBMCs) in promoting endothelial injury in r-axSpA. A total of 30 r-axSpA patients and 32 healthy donors (HDs) were evaluated. The endothelial function, inflammatory and atherogenic profile, and oxidative stress were quantified. In vitro studies were designed to evaluate the effect of PBMCs from r-axSpA patients on aberrant endothelial activation. Compared to HDs, our study found that, associated with ED and the plasma proatherogenic profile present in r-axSpA, PBMCs from these patients displayed a pro-oxidative, proinflammatory, and proatherogenic phenotype, with most molecular changes noticed in lymphocytes. Correlation studies revealed the relationship between this phenotype and the microvascular function. Additional in vitro studies confirmed that PBMCs from r-axSpA patients promoted endothelial injury. Altogether, this study suggests the relevance of r-axSpA itself as a strong and independent cardiovascular risk factor, contributing to a dysfunctional endothelium and atherogenic status by aberrant activation of PBMCs. Lymphocytes could be the main contributors in the development of ED and subsequent atherosclerosis in this pathology.

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