EOC 13.31 (ATCC® CRL-2468)

Organism: Mus musculus, mouse  /  Cell Type: microglia  /  Tissue: brain  / 

Organism Mus musculus, mouse
Tissue brain
Cell Type microglia
Product Format frozen
Morphology macrophage
Culture Properties adherent
Biosafety Level 1
Age 10 days
Gender female
Strain C3H/HeJ
Applications
The cells may be used to characterize the role of brain macrophages.
Storage Conditions liquid nitrogen vapor phase
Derivation

This is an immortalized cell line derived from the brain of an apparently normal 10 day old mouse. RefWalker WS, et al. Mouse microglial cell lines differing in constitutive and interferon-gamma-inducible antigen-presenting activities for naive and memory CD4+ and CD8+ T cells. J. Neuroimmunol. 63: 163-174, 1995. PubMed: 8550814 

Cells were cloned in soft agar in the presence of CSF-1 and expanded on microcarrier beads. Beads were transferred to culture dishes and were subsequently passaged by scraping.
Antigen Expression
CD11b/CD18 (Mac-1) +, Mac-2 +, Mac-3 +, CD80 (B7-1) +, CD86 (B7.2) +; CD45 +, Ly-6C +, F4/80 +, MHC Class I +, MHC Class II +, CD115 (colony stimulating factor 1 receptor (CSF-1R)) +, FcR +
Receptor Expression
colony stimulating factor 1 (CSF-1R, CD115)
Comments

The cell line is dependent on colony stimulating factor 1 (CSF-1).

The cells exhibit phagocytic activity.

These cells constitutively expressed high levels of major histocompatibility complex (MHC) class II antigens but unlike EOC-20 (CRL-2469) expression was not upregulated by recombinant murine interferon-gamma.

C3H/HeJ strain is defective in TLR4 (toll-like receptor 4)


Complete Growth Medium Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose, 70%; fetal bovine serum, 10%; LADMAC Conditioned Media (produced from the LADMAC cell line (CRL-2420), 20%
Subculturing
  1. Remove and discard 75% of the media.
  2. Scrape off the attached cells with a cell scraper.
  3. Add appropriate aliquots of cell suspension to new culture vessels.
  4. Incubate cultures at 37°C.

Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended
Medium Renewal: Every 2 to 3 days

Subculturing Procedure for LADMAC cells

Cultures can be established by centrifugation with subsequent resuspension at 2 X 10 5 viable cells/ml. Attached cells may be subcultured by tapping the sides of the flask until cells are dispersed.

Complete Growth Medium for LADMAC cells

The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003.  To make the complete growth medium, add the following components to the base medium: 

  • fetal bovine serum to a final concentration of 10%

This medium is formulated for use with a 5% CO2 in air atmosphere.

Cryopreservation Complete growth medium, 95%; DMSO, 5%. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.
Culture Conditions
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
Name of Depositor WS Walker
Passage History
Cells were cloned in soft agar in the presence of CSF-1 and expanded on microcarrier beads. Beads were transferred to culture dishes and were subsequently passaged by scraping.
References

Olivas E, et al. Use of the Pannell-Milstein roller bottle apparatus to produce high concentrations of the CSF-1, the mouse macrophage growth factor. J. Immunol. Methods 182: 73-79, 1995. PubMed: 7769247

Walker WS. Establishment of mononuclear phagocyte cell lines. J. Immunol. Methods 174: 25-31, 1994. PubMed: 8083530

Walker WS, et al. Mouse microglial cell lines differing in constitutive and interferon-gamma-inducible antigen-presenting activities for naive and memory CD4+ and CD8+ T cells. J. Neuroimmunol. 63: 163-174, 1995. PubMed: 8550814

Askew D, Walker WS. Alloantigen presentation to naive CD8+ T cells by mouse microglia: evidence for a distinct phenotype based on expression of surface-associated and soluble costimulatory molecules. Glia 18: 118-128, 1996. PubMed: 8913775

Basic Documentation
References

Olivas E, et al. Use of the Pannell-Milstein roller bottle apparatus to produce high concentrations of the CSF-1, the mouse macrophage growth factor. J. Immunol. Methods 182: 73-79, 1995. PubMed: 7769247

Walker WS. Establishment of mononuclear phagocyte cell lines. J. Immunol. Methods 174: 25-31, 1994. PubMed: 8083530

Walker WS, et al. Mouse microglial cell lines differing in constitutive and interferon-gamma-inducible antigen-presenting activities for naive and memory CD4+ and CD8+ T cells. J. Neuroimmunol. 63: 163-174, 1995. PubMed: 8550814

Askew D, Walker WS. Alloantigen presentation to naive CD8+ T cells by mouse microglia: evidence for a distinct phenotype based on expression of surface-associated and soluble costimulatory molecules. Glia 18: 118-128, 1996. PubMed: 8913775