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ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.
ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
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: 8550814Cells 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.
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)
To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C. Storage at -70°C will result in loss of viability.
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:
This medium is formulated for use with a 5% CO2 in air atmosphere.
The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid. Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.
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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