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'''Bone-marrow-derived [[macrophage]]''' ('''BMDM''') refers to [[macrophage]] cells that are generated in a research laboratory from mammalian [[bone marrow]] cells.<ref>{{cite journal | vauthors = Barrett JP, Costello DA, O'Sullivan J, Cowley TR, Lynch MA | title = Bone marrow-derived macrophages from aged rats are more responsive to inflammatory stimuli | journal = Journal of Neuroinflammation | volume = 12 | issue = 1 | pages = 67 | date = April 2015 | pmid = 25890218 | pmc = 4397943 | doi = 10.1186/s12974-015-0287-7 }}</ref> Undifferentiated bone marrow cells are cultured in the presence of [[macrophage colony-stimulating factor]] (M-CSF; CSF1).<ref>{{cite journal | vauthors = Weischenfeldt J, Porse B | title = Bone Marrow-Derived Macrophages (BMM): Isolation and Applications | journal = CSH Protocols | volume = 2008 | issue = 12 | pages = pdb.prot5080 | date = December 2008 | pmid = 21356739 | doi = 10.1101/pdb.prot5080 }}</ref> M-CSF is a [[cytokine]] that directs [[cell differentiation]] toward an M2 polarization. These cells are often used in [[immunology]] and [[cell biology]] research.<ref name="pmid15845473">{{cite journal | vauthors = Yoshizawa S, Tateda K, Matsumoto T, Gondaira F, Miyazaki S, Standiford TJ, Yamaguchi K | title = Legionella pneumophila evades gamma interferon-mediated growth suppression through interleukin-10 induction in bone marrow-derived macrophages | journal = Infection and Immunity | volume = 73 | issue = 5 | pages = 2709–17 | date = May 2005 | pmid = 15845473 | pmc = 1087334 | doi = 10.1128/IAI.73.5.2709-2717.2005 }}</ref>
'''Bone-marrow-derived [[macrophage]]''' ('''BMDM''') refers to [[macrophage]] cells that are generated in a research laboratory from mammalian [[bone marrow]] cells.<ref name=":0">{{cite journal|vauthors=Barrett JP, Costello DA, O'Sullivan J, Cowley TR, Lynch MA|date=April 2015|title=Bone marrow-derived macrophages from aged rats are more responsive to inflammatory stimuli|journal=Journal of Neuroinflammation|volume=12|issue=1|pages=67|doi=10.1186/s12974-015-0287-7|pmc=4397943|pmid=25890218}}</ref><ref name=":1">{{Cite journal|last=Li|first=Yue|last2=Niu|first2=Shixian|last3=Xi|first3=Dalin|last4=Zhao|first4=Shuqi|last5=Sun|first5=Jiang|last6=Jiang|first6=Yong|last7=Liu|first7=Jinghua|date=2019|title=Differences in Lipopolysaccharides-Induced Inflammatory Response Between Mouse Embryonic Fibroblasts and Bone Marrow-Derived Macrophages|url=https://www.ncbi.nlm.nih.gov/pubmed/30990360|journal=Journal of Interferon & Cytokine Research: The Official Journal of the International Society for Interferon and Cytokine Research|volume=39|issue=6|pages=375–382|doi=10.1089/jir.2018.0167|issn=1557-7465|pmid=30990360|via=}}</ref><ref name=":2" /> BMDMs can differentiate into mature macrophages in the presence of [[Growth factor|growth factors]] and other signaling molecules.<ref name=":0" /><ref name=":1" /> Undifferentiated bone marrow cells are cultured in the presence of [[macrophage colony-stimulating factor]] (M-CSF; CSF-1).<ref name=":2">{{cite journal | vauthors = Weischenfeldt J, Porse B | title = Bone Marrow-Derived Macrophages (BMM): Isolation and Applications | journal = CSH Protocols | volume = 2008 | issue = 12 | pages = pdb.prot5080 | date = December 2008 | pmid = 21356739 | doi = 10.1101/pdb.prot5080 }}</ref> M-CSF is a [[cytokine]] and growth factor that is responsible for the proliferation and commitment of [[Myeloid tissue|myeloid]] [[Progenitor|progenitors]] into [[Monocyte|monocytes]] (which then mature into macrophages).<ref name=":2" /><ref>{{Cite journal|last=Hamilton|first=Thomas A.|last2=Zhao|first2=Chenyang|last3=Pavicic|first3=Paul G.|last4=Datta|first4=Shyamasree|date=2014-11-21|title=Myeloid Colony-Stimulating Factors as Regulators of Macrophage Polarization|url=https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4240161/|journal=Frontiers in Immunology|volume=5|doi=10.3389/fimmu.2014.00554|issn=1664-3224|pmc=4240161|pmid=25484881}}</ref> Macrophages have a wide variety of functions in the body including [[phagocytosis]] of foreign invaders and other cellular debris, cytokine release to trigger immune responses, and antigen presentation.<ref name=":1" /> BMDMs provide a large homogenous population of macrophages that play an increasingly important role in making macrophage related research possible and financially feasible.<ref name=":3">{{Cite journal|last=Marim|first=Fernanda M.|last2=Silveira|first2=Tatiana N.|last3=Lima|first3=Djalma S.|last4=Zamboni|first4=Dario S.|date=2010-12-17|title=A method for generation of bone marrow-derived macrophages from cryopreserved mouse bone marrow cells|url=https://www.ncbi.nlm.nih.gov/pubmed/21179419|journal=PloS One|volume=5|issue=12|pages=e15263|doi=10.1371/journal.pone.0015263|issn=1932-6203|pmc=3003694|pmid=21179419}}</ref>

== Production ==
[[File:BMDM_production.png|thumb|Schema of in vitro BMDM production]]
In order to produce BMDMs, [[Mesenchymal stem cell|mesenchymal stem cells]] are removed from tibia or femur of mice.<ref name=":4">{{Cite journal|last=Hume|first=D. A.|last2=Allan|first2=W.|last3=Fabrus|first3=B.|last4=Weidemann|first4=M. J.|last5=Hapel|first5=A. J.|last6=Bartelmez|first6=S.|date=1987|title=Regulation of proliferation of bone marrow-derived macrophages|url=https://www.ncbi.nlm.nih.gov/pubmed/3035291|journal=Lymphokine Research|volume=6|issue=2|pages=127–139|issn=0277-6766|pmid=3035291}}</ref> Since BMDMs are derived from bone marrow, derived cells are healthy and naïve (or unactivated), regardless of the condition of donor mice.<ref name=":3" /> The femoral or tibia bone marrow cells are then incubated with CSF-1.<ref name=":4" /> Without CSF-1, the cells enter an inactive state but can reinitiate DNA synthesis if stimulated later.<ref name=":4" /> Mature macrophages and fibroblasts, which may carry unwanted growth factors, are removed. [[Interleukin 3|IL-3]] and [[Interleukin-1 family|IL-1]], two growth factors, are often added to increase yield and promote rapid terminal differentiation.<ref name=":4" /> Exogenous media containing growth factors and other serums must also be added to make the cells continually viable.<ref name=":4" /> Full growth and differentiation take approximately 5-8 days.<ref name=":4" /> Millions of BMDMs can be derived from one mouse; cells are then frozen for years and can be thawed and respond to a variety of stimuli such as [[Lipopolysaccharide|LPS]], [[Interferon gamma|IFN-γ]], [[Pathogen-associated molecular pattern|PAMPs]], [[NF-κB]], and [[IRF3]].<ref name=":0" /><ref name=":3" /><ref>{{Cite journal|last=Oppong-Nonterah|first=Gertrude O.|last2=Lakhdari|first2=Omar|last3=Yamamura|first3=Asami|last4=Hoffman|first4=Hal M.|last5=Prince|first5=Lawrence S.|date=2019|title=TLR Activation Alters Bone Marrow-Derived Macrophage Differentiation|url=https://www.ncbi.nlm.nih.gov/pubmed/30408777|journal=Journal of Innate Immunity|volume=11|issue=1|pages=99–108|doi=10.1159/000494070|issn=1662-8128|pmc=6296861|pmid=30408777}}</ref> These signals induce translation of genes that produce cytokines and determine if macrophages are M1 (pro-inflammatory) or M2 (anti-inflammatory).<ref name=":1" /> If BMDMs are not frozen, they become less viable with age as CSF-1 concentration decreases.<ref name=":0" />

Proliferation of BMDMs can also be inhibited by a number of reagents.<ref name=":4" /> For example, growth and differentiation is dependent on CSF-1 and a functional CSF-1 receptor, a member of the [[tyrosine kinase]] family, on stem cells.<ref name=":4" /> [[Interferon|Interferons]] can cause a down regulation of CSF-1 receptor.<ref name=":4" /> Additionally, without stimuli like LPS to induce macrophage maturation to M1 or M2, mice accumulate immature macrophages which are less helpful to the body.<ref name=":4" />
<br />
== References ==
== References ==
{{Reflist}}
{{Reflist}}

Revision as of 00:47, 10 March 2020

Bone-marrow-derived macrophage (BMDM) refers to macrophage cells that are generated in a research laboratory from mammalian bone marrow cells.[1][2][3] BMDMs can differentiate into mature macrophages in the presence of growth factors and other signaling molecules.[1][2] Undifferentiated bone marrow cells are cultured in the presence of macrophage colony-stimulating factor (M-CSF; CSF-1).[3] M-CSF is a cytokine and growth factor that is responsible for the proliferation and commitment of myeloid progenitors into monocytes (which then mature into macrophages).[3][4] Macrophages have a wide variety of functions in the body including phagocytosis of foreign invaders and other cellular debris, cytokine release to trigger immune responses, and antigen presentation.[2] BMDMs provide a large homogenous population of macrophages that play an increasingly important role in making macrophage related research possible and financially feasible.[5]

Production

Schema of in vitro BMDM production

In order to produce BMDMs, mesenchymal stem cells are removed from tibia or femur of mice.[6] Since BMDMs are derived from bone marrow, derived cells are healthy and naïve (or unactivated), regardless of the condition of donor mice.[5] The femoral or tibia bone marrow cells are then incubated with CSF-1.[6] Without CSF-1, the cells enter an inactive state but can reinitiate DNA synthesis if stimulated later.[6] Mature macrophages and fibroblasts, which may carry unwanted growth factors, are removed. IL-3 and IL-1, two growth factors, are often added to increase yield and promote rapid terminal differentiation.[6] Exogenous media containing growth factors and other serums must also be added to make the cells continually viable.[6] Full growth and differentiation take approximately 5-8 days.[6] Millions of BMDMs can be derived from one mouse; cells are then frozen for years and can be thawed and respond to a variety of stimuli such as LPS, IFN-γ, PAMPs, NF-κB, and IRF3.[1][5][7] These signals induce translation of genes that produce cytokines and determine if macrophages are M1 (pro-inflammatory) or M2 (anti-inflammatory).[2] If BMDMs are not frozen, they become less viable with age as CSF-1 concentration decreases.[1]

Proliferation of BMDMs can also be inhibited by a number of reagents.[6] For example, growth and differentiation is dependent on CSF-1 and a functional CSF-1 receptor, a member of the tyrosine kinase family, on stem cells.[6] Interferons can cause a down regulation of CSF-1 receptor.[6] Additionally, without stimuli like LPS to induce macrophage maturation to M1 or M2, mice accumulate immature macrophages which are less helpful to the body.[6]

References

  1. ^ a b c d Barrett JP, Costello DA, O'Sullivan J, Cowley TR, Lynch MA (April 2015). "Bone marrow-derived macrophages from aged rats are more responsive to inflammatory stimuli". Journal of Neuroinflammation. 12 (1): 67. doi:10.1186/s12974-015-0287-7. PMC 4397943. PMID 25890218.{{cite journal}}: CS1 maint: unflagged free DOI (link)
  2. ^ a b c d Li, Yue; Niu, Shixian; Xi, Dalin; Zhao, Shuqi; Sun, Jiang; Jiang, Yong; Liu, Jinghua (2019). "Differences in Lipopolysaccharides-Induced Inflammatory Response Between Mouse Embryonic Fibroblasts and Bone Marrow-Derived Macrophages". Journal of Interferon & Cytokine Research: The Official Journal of the International Society for Interferon and Cytokine Research. 39 (6): 375–382. doi:10.1089/jir.2018.0167. ISSN 1557-7465. PMID 30990360.
  3. ^ a b c Weischenfeldt J, Porse B (December 2008). "Bone Marrow-Derived Macrophages (BMM): Isolation and Applications". CSH Protocols. 2008 (12): pdb.prot5080. doi:10.1101/pdb.prot5080. PMID 21356739.
  4. ^ Hamilton, Thomas A.; Zhao, Chenyang; Pavicic, Paul G.; Datta, Shyamasree (2014-11-21). "Myeloid Colony-Stimulating Factors as Regulators of Macrophage Polarization". Frontiers in Immunology. 5. doi:10.3389/fimmu.2014.00554. ISSN 1664-3224. PMC 4240161. PMID 25484881.{{cite journal}}: CS1 maint: unflagged free DOI (link)
  5. ^ a b c Marim, Fernanda M.; Silveira, Tatiana N.; Lima, Djalma S.; Zamboni, Dario S. (2010-12-17). "A method for generation of bone marrow-derived macrophages from cryopreserved mouse bone marrow cells". PloS One. 5 (12): e15263. doi:10.1371/journal.pone.0015263. ISSN 1932-6203. PMC 3003694. PMID 21179419.{{cite journal}}: CS1 maint: unflagged free DOI (link)
  6. ^ a b c d e f g h i j Hume, D. A.; Allan, W.; Fabrus, B.; Weidemann, M. J.; Hapel, A. J.; Bartelmez, S. (1987). "Regulation of proliferation of bone marrow-derived macrophages". Lymphokine Research. 6 (2): 127–139. ISSN 0277-6766. PMID 3035291.
  7. ^ Oppong-Nonterah, Gertrude O.; Lakhdari, Omar; Yamamura, Asami; Hoffman, Hal M.; Prince, Lawrence S. (2019). "TLR Activation Alters Bone Marrow-Derived Macrophage Differentiation". Journal of Innate Immunity. 11 (1): 99–108. doi:10.1159/000494070. ISSN 1662-8128. PMC 6296861. PMID 30408777.
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