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Immortalized Mouse Microglia (SIM-A9)

SKU : BHC10900793

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Applied Biological Materials (abm) Inc.

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Mfr.No.	T0247
Description	Stimulation of microglia can be done by adding 2.5 ng/ml LPS, or 20 ?M ?-amyloid fibrils (A?1-42); 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 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 ?-amyloid) and the ability to secrete 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
Biosafety Level	2
Depositor	University of North Dakota
Disclaimer	1. 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 ?g, Cat.# C207, $450.00) or cell lysate (100 ?g, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 2. 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). 3. 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. 4. 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. 5. 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."
Donor History	C57BL/6 mouse
Expression Region	CD68, Iba1
Growth Properties	Adherent, round or polygonal
Growth Conditions	PriCoat?T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) are recommended for optimal cell culture.?riGrow IV (TM004) + 10% heat-inactivated FBS + 5% horse serum + 1% Penicillin/Streptomycin Solution (G255), 37.0?, 5% CO?.
Quantity	1x106 cells / 1.0 ml
Tissue	Brain
Morphology	Cobble-stone
Notes	1. Visually examine the packaging containers for signs of leakage or breakage. 2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130?, preferably in liquid nitrogen vapor phase storage, until ready for use. To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130? or in liquid nitrogen vapor phase. Do not store at -70?, as it will result in loss of viability.
Organism	Mouse (M. musculus)
Species	Mouse (M. musculus)
Propagation	Use of PriCoat T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Thaw the cells with PriGrow XI (TM011) + 8% FBS + 60 µM CaCl? (added fresh each time) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 8% CO? for optimal attachment. After 24h, change media to PriGrow XI (TM011) + 8% chelexed FBS + 1.4% FBS + 5 ng/ml rhFGF7 (Z100595) (freshly added) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 8% CO?Steps to thaw the cells:1. Thaw the cells in RT water until only a few ice crystals are left2. Put the cells in 5 ml of RT PriGrow XI (TM011) + 8% FBS3. Centrifuge the cells 1500 rpm for 5 min (need this step to remove inhibitors for attachment)4. Resuspend the pellet in RT Prigrow XI (TM011) + 8% FBS + 1% P/S with freshly added 60 µM CaCl? and put the cells in a culture vessel at 37.0°C, 8% CO?5. After 18h-24h, change to PriGrow XI (TM011) + 8% chelexed FBS + 1.4% FBS + 5 ng/ml rhFGF7 (Z100595) (freshly added) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 8% CO? for growth of the cells.
Cryopreservation	Cryopreservation Medium (TM024), or complete growth media with 10% DMSO.
Quality Control	Immunoblot for the presence of CDC25A was conducted.

Applications Range	Research Use Only.

Shipping	Ship with dry ice.
Product Format	Frozen
Product Use	This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Storage	Vapor phase of liquid nitrogen, or below -130?.

Thawing Protocol	1. Thaw cells quickly in a 37? water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination. 2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions. 3. Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes. 4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask. 5. Incubate the cells at the recommended conditions.
Subculture Protocol	Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent. 1. Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel. 2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37?, for several minutes to facilitate detachment. 3. Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel. 4. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type. 5. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired. 6. Incubate the cells at the recommended conditions.

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SKU	Supplier No.	Size	Your price	Quantity
BHC10900793	T0247	1x10^6 cells / 1.0 ml	-		Add to Quote

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