Open AccessPMA withdrawal in PMA‐treated monocytic THP‐1 cells and subsequent retinoic acid stimulation, modulate induction of apoptosis and appearance of dendritic cells
Author(s) -
Spano A.,
Barni S.,
Sciola L.
Publication year - 2013
Publication title -
cell proliferation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.647
H-Index - 74
eISSN - 1365-2184
pISSN - 0960-7722
DOI - 10.1111/cpr.12030
Subject(s) - retinoic acid , cd14 , thp1 cell line , apoptosis , monocyte , flow cytometry , integrin alpha m , microbiology and biotechnology , phorbol , macrophage , cellular differentiation , dendritic cell , biology , tretinoin , cell culture , chemistry , immunology , in vitro , biochemistry , signal transduction , immune system , genetics , protein kinase c , gene
Objectives To analyse proliferation, differentiation and apoptosis in THP ‐1 cells after stimulation with phorbol 12‐myristate 13‐acetate ( PMA ) and retinoic acid ( RA ). Materials and methods PMA and RA were used in a three‐step‐procedure: (i) treatment with 6, 30, 60 n m PMA, that induced initial, intermediate and advanced levels of monocyte‐macrophage transition, respectively; (ii) recovery in PMA‐free medium; (iii) incubation with 4 μ m RA. Cultures were characterized cytokinetically (flow cytometry/bromodeoxyuridine uptake) and immunocytochemically (static cytometry) for expression of CD14, CD11b (monocyte‐macrophage) and DC‐SIGN (dendritic cell: DCs) markers. Results Some treatments determined appearance of monocyte/macrophage, dendritic and apoptotic phenotypes, percentages of which were related to PMA dose used in step 1, and dependent on presence/absence of PMA and RA. PMA withdrawal induced dedifferentiation and partial restoration of proliferative activity, specially in 6 and 30 n m PMA‐derived cells. Recovery in the presence of serum (fundamental to DC appearance) indicated that depending on differentiation level, cell proliferation and apoptosis were inversely correlated. Treatment with 30 n m PMA induced intermediate levels of monocytic‐macrophagic differentiation, with expression of alternative means of differentiation and acquisition of DCs without using cytokines, after PMA withdrawal and RA stimulation. Conclusions Our experimental conditions favoured differentiation, dedifferentiation and transdifferentiational pathways, in monocytic THP ‐1 cells, the balance of which could be related to both cell proliferation and cell death.