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...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2013-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3790761?pdf=render |
| _version_ | 1819128948504657920 |
|---|---|
| author | Sachiko Ito Yuriko Tanaka Naomi Nishio Suganya Thanasegaran Ken-Ichi Isobe |
| author_facet | Sachiko Ito Yuriko Tanaka Naomi Nishio Suganya Thanasegaran Ken-Ichi Isobe |
| author_sort | Sachiko Ito |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-22T08:35:56Z |
| format | Article |
| id | doaj.art-ec64c733d2c449449c97d67471060528 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-22T08:35:56Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-ec64c733d2c449449c97d674710605282022-12-21T18:32:21ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-01810e7694310.1371/journal.pone.0076943Establishment of self-renewable GM-CSF-dependent immature macrophages in vitro from murine bone marrow.Sachiko ItoYuriko TanakaNaomi NishioSuganya ThanasegaranKen-Ichi IsobeMacrophages 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.http://europepmc.org/articles/PMC3790761?pdf=render |
| spellingShingle | Sachiko Ito Yuriko Tanaka Naomi Nishio Suganya Thanasegaran Ken-Ichi Isobe Establishment of self-renewable GM-CSF-dependent immature macrophages in vitro from murine bone marrow. PLoS ONE |
| title | Establishment of self-renewable GM-CSF-dependent immature macrophages in vitro from murine bone marrow. |
| title_full | Establishment of self-renewable GM-CSF-dependent immature macrophages in vitro from murine bone marrow. |
| title_fullStr | Establishment of self-renewable GM-CSF-dependent immature macrophages in vitro from murine bone marrow. |
| title_full_unstemmed | Establishment of self-renewable GM-CSF-dependent immature macrophages in vitro from murine bone marrow. |
| title_short | Establishment of self-renewable GM-CSF-dependent immature macrophages in vitro from murine bone marrow. |
| title_sort | establishment of self renewable gm csf dependent immature macrophages in vitro from murine bone marrow |
| url | http://europepmc.org/articles/PMC3790761?pdf=render |
| work_keys_str_mv | AT sachikoito establishmentofselfrenewablegmcsfdependentimmaturemacrophagesinvitrofrommurinebonemarrow AT yurikotanaka establishmentofselfrenewablegmcsfdependentimmaturemacrophagesinvitrofrommurinebonemarrow AT naominishio establishmentofselfrenewablegmcsfdependentimmaturemacrophagesinvitrofrommurinebonemarrow AT suganyathanasegaran establishmentofselfrenewablegmcsfdependentimmaturemacrophagesinvitrofrommurinebonemarrow AT kenichiisobe establishmentofselfrenewablegmcsfdependentimmaturemacrophagesinvitrofrommurinebonemarrow |