PMID- 23895502
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20140520
LR  - 20240321
IS  - 2045-3701 (Print)
IS  - 2045-3701 (Electronic)
IS  - 2045-3701 (Linking)
VI  - 3
DP  - 2013
TI  - Cell density during differentiation can alter the phenotype of bone 
      marrow-derived macrophages.
PG  - 30
LID - 10.1186/2045-3701-3-30 [doi]
AB  - BACKGROUND: Bone marrow-derived macrophages (BMDMs) are widely used primary cells 
      for studying macrophage function. However, despite numerous protocols that are 
      currently available, lack of a notable consensus on generating BMDMs may obscure 
      the reliability in comparing findings from different studies or laboratories. 
      FINDINGS: In this study, we addressed the effect of cell density on the resulting 
      macrophage population. With reference to previously published methods, bone 
      marrow cells from wild type C57BL/6 mice were plated at either 4 x 10(5) cells or 
      5 x 10(6) cells per 10 cm and cultured in 20% L-cell conditioned media for 
      7 days, after which they were analyzed for cell surface markers, production of 
      proinflammatory cytokines, and responsiveness to polarizing signals. 
      Reproducibly, cells plated at lower density gave a pure population of 
      CD11b(+)F4/80(+) macrophages (97.28 +/- 0.52%) with majority being Ly-6C(-)Ly-6G(-) 
      and c-Fms(+), while those plated at higher density produced less CD11b(+)F4/80(+) 
      cells and a considerably higher proportion of CD11b(+)F4/80(+)CD11c(+) 
      (68.72 +/- 2.52%) and Ly-6C(-)Ly-6G(+) (71.10 +/- 0.90%) cells. BMDMs derived from 
      higher plating density also secreted less proinflammatory cytokines such as IL-6, 
      IL-12 and TNF-alpha and were less phagocytic, and had a different pattern of 
      expression for M1- and M2-related genes upon LPS or IL-4 stimulation. 
      CONCLUSIONS: Overall, our findings indicate that altering cell density during 
      BMDM differentiation can give rise to distinct macrophage populations that could 
      vary the outcome of a functional study.
FAU - Lee, Chan Mi
AU  - Lee CM
AD  - Physiology and Experimental Medicine, SickKids, 555 University Avenue, M5G 1X8, 
      Toronto, Ontario, Canada ; Laboratory Medicine & Pathobiology, University of 
      Toronto, 1 King's College Circle, M5S 1A8, Toronto, Ontario, Canada.
FAU - Hu, Jim
AU  - Hu J
AD  - Physiology and Experimental Medicine, SickKids, 555 University Avenue, M5G 1X8, 
      Toronto, Ontario, Canada ; Laboratory Medicine & Pathobiology, University of 
      Toronto, 1 King's College Circle, M5S 1A8, Toronto, Ontario, Canada.
LA  - eng
PT  - Journal Article
DEP - 20130729
PL  - England
TA  - Cell Biosci
JT  - Cell & bioscience
JID - 101561195
PMC - PMC3750618
OTO - NOTNLM
OT  - Bone marrow-derived macrophages
OT  - Macrophage phenotype
OT  - Plating density
EDAT- 2013/07/31 06:00
MHDA- 2013/07/31 06:01
PMCR- 2013/07/29
CRDT- 2013/07/31 06:00
PHST- 2013/05/31 00:00 [received]
PHST- 2013/07/10 00:00 [accepted]
PHST- 2013/07/31 06:00 [entrez]
PHST- 2013/07/31 06:00 [pubmed]
PHST- 2013/07/31 06:01 [medline]
PHST- 2013/07/29 00:00 [pmc-release]
AID - 2045-3701-3-30 [pii]
AID - 10.1186/2045-3701-3-30 [doi]
PST - epublish
SO  - Cell Biosci. 2013 Jul 29;3:30. doi: 10.1186/2045-3701-3-30. eCollection 2013.