Microglia cells are resident macrophages of the brain and spinal cord and they act as the first and the main form of active immune defense in the nervous system. In addition to expressing microglial specific markers CD68 and Iba1, the spontaneously Immortalized Mouse Microglia (SIM-A9) retains responsiveness to exogenous inflammatory stimulation (LPS and beta-amyloid) and the ability to secret cytokines and nitric oxide as well as phagocytose biological debris. Upon LPS and IL-4 stimulation, these cells are capable of switching between the pro-inflammatory microglial phenotype, M1, which is associated with elevated iNOS and COX-2 followed by I?B and tyrosine kinase pathways and the regenerative-supportive phenotype, M2, which is identified by an increased in Arg-1 expression, respectively. SIM-A9 exhibits key attributes of primary microglia and is useful in characterization of stimulus-triggered microglial migration, proliferation and phagocytosis.
Cell Type
Immortalized Cells
Disclaimer
1. For for-profit organizations and corporations, please contact [email protected] for pricing of this item. 2. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each order. If you have any questions regarding this, please contact us at [email protected]. 3. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 mug, Cat.# C207, $450.00) or cell lysate (100 mug, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 4. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping). 5. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information. 6. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. 7. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period."
Expression Level
CD68, Iba1
Growth Properties
Adherent
Quantity
1x106 cells / 1.0 ml
Tissue
Brain
Morphology
Round or Polygonal
Population Doubling
30 - 40 hours
Species
Mouse (M. musculus)
Propagation
Use of PriCoatTMT25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells inECM-coated culture vessels unless otherwise specified in the Propagation Requirements below.The base medium for this cell line is Prigrow IV medium available at abm (TM004). To make the complete growth medium, add the following components to the base medium: heat-inactivated fetal bovine serum to a final concentration of 10% donor horse serum (Thermo Fisher Scientific; Cat. no. 1605013) to a final concentratio of 5% and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%.Change media every 2-3 days.Carbon dioxide (CO2): 5%, Temperature: 37.0°C.Stimulation of microglia can be done by adding 2.5 ng/ml LPS, or 20 muM beta-amyloid fibrils (Abeta1-42).
Quality Control
1) Functional assays such as the nitrate assay was performed to assess nitric oxide production; 2) ELISA was used to determine the TNFalpha secretion after inflammatory stimulation; 3) Western blot and immunocytochemistry were used to confirm the expression of microglial specific markers such as CD68 and Iba1 and M1/M2 phenotype-associated protein [removed]COX-2/iNOS and Arg-1) after stimulation; 4) Abeta1-42 or E. coli-derived bioparticles uptake was used to determine phagocytic activity of SIM-A9 cells.
Reviews of Immortalized Mouse Mesangial Cells - SV40