Establishment of self-renewable GM-CSF-dependent immature macrophages in vitro from murine bone marrow

Macrophages play a key role in the innate immune system. Macrophages are thought to originate from hematopoietic precursors or the yolk sac. Here, we describe the in vitro establishment of self-renewable GM-CSF-dependent immature macrophages (GM-IMs) from murine bone marrow (BM). GM-IMs grow continu...

Full description

Saved in:
Bibliographic Details
Published in:PloS one 2013-10, Vol.8 (10), p.e76943-e76943
Main Authors: Ito, Sachiko, Tanaka, Yuriko, Nishio, Naomi, Thanasegaran, Suganya, Isobe, Ken-Ichi
Format: Article
Language:English
Subjects:
Animals
Bisphosphonates
Bone marrow
Bone Marrow Cells - cytology
Bone Marrow Cells - drug effects
Bone Marrow Cells - metabolism
CD11b antigen
Cell culture
Cell cycle
Cell Differentiation
Cell Lineage
Cell Proliferation - drug effects
Chemokines
Clodronic acid
Conditioning
Cytokines
Dendritic cells
Female
Gene expression
Gene Expression Profiling
Granulocyte-macrophage colony-stimulating factor
Granulocyte-Macrophage Colony-Stimulating Factor - metabolism
Granulocyte-Macrophage Colony-Stimulating Factor - pharmacology
Hematopoietic Stem Cell Transplantation
Hemopoiesis
Immune system
Immunology
Immunophenotyping
Inflammation
Innate immunity
Interleukin 13
Interleukin 4
Interleukin 6
Lipopolysaccharides
Lymphocytes T
Macrophage colony-stimulating factor
Macrophages
Macrophages - cytology
Macrophages - drug effects
Macrophages - metabolism
Markers
Medicine
Mice
Monocytes
Myeloid Progenitor Cells - cytology
Myeloid Progenitor Cells - drug effects
Myeloid Progenitor Cells - metabolism
Phagocytes
Phenotype
Precursors
Primary Cell Culture
Proteins
Receptors
Rodents
Science
Stem cells
Stimulation
Tumor necrosis factor-α
University graduates
Wound healing
Wounding
Yolk
γ-Interferon
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Macrophages play a key role in the innate immune system. Macrophages are thought to originate from hematopoietic precursors or the yolk sac. Here, we describe the in vitro establishment of self-renewable GM-CSF-dependent immature macrophages (GM-IMs) from murine bone marrow (BM). GM-IMs grow continuously in vitro in conditioned medium containing GM-CSF. The immunophenotype of GM-IMs is F4/80(high) CD11b(high) CD11c(low) Ly6C(low). By comparing gene expression in GM-IMs and BM dendritic cells, we found that GM-IMs expressed lower levels of chemokines, cytokines and their receptors. GM-IMs are round in shape, attach loosely to non-coated culture dishes and have a marked phagocytic capacity. These results indicate that GM-IMs are macrophage precursor cells. Following stimulation with LPS, monocyte-like GM-IMs converted to flat macrophage-like cells that tightly adhered to non-coated culture dishes and produced pro-inflammatory cytokines TNFα, IL-6 and IL-1β. These results indicated that GM-IMs differentiated to M1 pro-inflammatory macrophages. This was confirmed by stimulation of GM-IMs with IFNγ, an inducer of M1 markers. GM-IMs showed enhanced expression of M2 macrophage markers such as Arg1 and Retnla following stimulation by Th2 cytokines IL-4 and IL-13. When GM-IMs were injected into mice at sites of wounding, wound repair was enhanced. These results indicate that GM-IMs can differentiate to M2 macrophages. When GM-IMs were injected into clodronate-treated mice, they induced resident macrophage proliferation by producing M-CSF. In conclusion we have established self-renewable GM-CSF-dependent immature macrophages in vitro from murine BM, which differentiate to M1 or M2 macrophages.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0076943