Culturing cells with mast cell phenotype and function: Comparison of peripheral blood and bone marrow as a source.
Biomarkers
/ metabolism
Biopsy
Bone Marrow Cells
/ immunology
Bone Marrow Examination
Case-Control Studies
Cell Culture Techniques
Cell Degranulation
Cell Separation
Cells, Cultured
Flow Cytometry
Humans
Immunophenotyping
Mast Cells
/ immunology
Mastocytosis, Systemic
/ diagnosis
Phenotype
Time Factors
CD117
CD63
Flow cytometry
Human mast cell
MRGPRX2
Mast cell activation
Substance P
Journal
Journal of immunological methods
ISSN: 1872-7905
Titre abrégé: J Immunol Methods
Pays: Netherlands
ID NLM: 1305440
Informations de publication
Date de publication:
08 2021
08 2021
Historique:
received:
12
02
2021
revised:
17
04
2021
accepted:
26
04
2021
pubmed:
3
5
2021
medline:
26
10
2021
entrez:
2
5
2021
Statut:
ppublish
Résumé
Studies on the mechanisms that govern mast cell (MC) functions are hindered by the difficulties in isolating sufficient numbers of these tissue-resident cells. Therefore, many research groups use cultured human MCs obtained out of progenitor cells. However, these culture methods significantly differ regarding primary source material, culture durations and conditions. Consequently, the finally obtained cells are likely to exhibit morphological, phenotypical and/or functional heterogeneity. To compare the phenotype and functionality of cells cultured from peripheral blood and bone marrow progenitor cells from patients with suspected clonal MC disease. These cells are designated as PBCMCs and BMCMCs, respectively. Twenty paired PBCMCs and BMCMCs cultures starting from CD34 PBCMCs and BMCMCs are phenotypically comparable. Functionally, after activation with anti-FcεRI and substance P, PBCMCs and BMCMCs show similar up-regulation of the lysosomal degranulation marker CD63. However, the yield of PBCMCs is higher than BMCMs and peripheral blood cultures are purer than bone marrow cultures. PBCMCs are an attractive alternative to the more difficult to obtain BMCMCs for the exploration of the complex mechanisms that govern IgE- and MRGPRX2-dependent MC activation and degranulation. Unlike BMCMCs, PBCMCs are easily accessible and enable repetitive analyses.
Sections du résumé
BACKGROUND
Studies on the mechanisms that govern mast cell (MC) functions are hindered by the difficulties in isolating sufficient numbers of these tissue-resident cells. Therefore, many research groups use cultured human MCs obtained out of progenitor cells. However, these culture methods significantly differ regarding primary source material, culture durations and conditions. Consequently, the finally obtained cells are likely to exhibit morphological, phenotypical and/or functional heterogeneity.
OBJECTIVE
To compare the phenotype and functionality of cells cultured from peripheral blood and bone marrow progenitor cells from patients with suspected clonal MC disease. These cells are designated as PBCMCs and BMCMCs, respectively.
METHODS
Twenty paired PBCMCs and BMCMCs cultures starting from CD34
RESULTS
PBCMCs and BMCMCs are phenotypically comparable. Functionally, after activation with anti-FcεRI and substance P, PBCMCs and BMCMCs show similar up-regulation of the lysosomal degranulation marker CD63. However, the yield of PBCMCs is higher than BMCMs and peripheral blood cultures are purer than bone marrow cultures.
CONCLUSION
PBCMCs are an attractive alternative to the more difficult to obtain BMCMCs for the exploration of the complex mechanisms that govern IgE- and MRGPRX2-dependent MC activation and degranulation. Unlike BMCMCs, PBCMCs are easily accessible and enable repetitive analyses.
Identifiants
pubmed: 33933470
pii: S0022-1759(21)00106-X
doi: 10.1016/j.jim.2021.113061
pii:
doi:
Substances chimiques
Biomarkers
0
Types de publication
Comparative Study
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
113061Informations de copyright
Copyright © 2021 Elsevier B.V. All rights reserved.