Immortalized Mouse Dendritic Cells (MutuDC1940)

Mfr.No.	T0528
Description	As messengers bridging the innate and adaptive immune systems, conventional/classical lymphoid tissue-resident dendritic cells (cDCs) capture and present antigens; acting as sentinels in secondary lymphoid organs and other tissues. The wild-type MutuDC1940 dendritic cell line is derived from mouse spleen tissues (CD11c:SV40LgT transgenic mice) and is a powerful tool for vaccine science and immunotherapy, particularly for strategizing target antigens to the CD8?+?ubset (which has the CD11chigh, B220?, DEC205+, CD24high, CD11b??xpression profile).?br />MutuDC cell lines are GFP positive due to the GFP reporter in the CD11c:SV40LgT transgene. This cell line has the following characteristics:responds to TLR ligands such as CpG (TLR9-L), PolyIC (TLR3-L), and LPS (TLR4-L) by up-regulation of CD40, CD80 and CD86responds to PAMP stimulation by production of Th1 cytokines such as IL-12capable of MHC-I and MHC-II antigen-presentation (both direct and cross-presentation of cell-associated antigens; evaluated by by the MHC-I (SIINFEKL/OT-I ) and MHC-II (OVA323-339/OT-II)?estricted systems) Additional MutuDC cell lines available at abm include:Cat. T3034 - TLR3 Knockout MutuDC Cell line?at. T3035 - TLR9 Knockout MutuDC Cell line?at.?3036 - Ifnar1 Knockout MutuDC Cell line
Cell Type	Immortalized Cells
Biosafety Level	2
Depositor	University of Lausanne (PACTT)
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	CD11c:SV40LgT-transgenic C57BL/6 mice
Expression Region	GFP, CD11c, CD24, MHC-II+, DEC205, Clec9A, B220, CD11b, IRF4, IRF8, CD4
Expression Level	CD73, CD44, CD90/Thy-1, CD105/Endoglin, CD166/ALCAM, CD133/Prom1, BMPRII, CD117/c-kit, and CD29/integrin beta1
Growth Properties	Adherent, small aggregates
Growth Conditions	Use of PriCoat?T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is recommended for cell adhesion to the culture vessels. Prigrow V (TM015) + 4 mM Glutamax + heat-inactivated 10% FBS + 1% of 7.5% Sodium Bicarbonate Solution + 50 ? ?-mercaptoethanol + 10 mM ?EPES (TM058) + 1% Penicillin/Streptomycin Solution (G255), 37.0?, 5% CO?.?ilter the complete media with a 0.22? filter prior to use.Note: cells must be seeded and maintained at high density for optimal growth. Cells are sensitive to culture conditions; change the complete media every 2-3 days.?o not let media colour change to yellow.
Quantity	1x106 cells / 1.0 ml
Tissue	Spleen
Morphology	Polygonal
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 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 III medium available at abm, Cat. No. TM003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 10% 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.* Do not use heat-inactivated FBS for cell culture unless specified otherwise.
Cryopreservation	Cryopreservation Medium (TM024), or complete growth media + 50% HI-FBS + 10% DMSO.
Quality Control	1) Immunofluorescence staining performed to examine mesenchymal stem cell markers; 2) sqPCR analysis for BMP9-induced late osteo/odontoblast, and ondotoblast markers.

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 290xg for 5 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 85-90% confluent.?se a prewarmed (37?)?on-enzymatic, 5?mM EDTA-based cell dissociation buffer (5?mM EDTA in 20?mM Hepes-PBS) to subculture cells. 1. Aspirate excess media, gently wash cells with PBS (1X) and add?-3 ml of the pre-warmed (37?) disssociation buffer mixture to the culture vessel.?br>2. Incubate culture vessel at 37? for 3-5 minutes to facilitate detachment.?bserve the cells under a microscope to confirm detachment. Gently tap the flask to dislodge any loosely adherent cells.?br>3. Neutralize the solution 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 290xg 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. Cells must be seeded at high density.?br>6. Incubate the cells at the recommended conditions.

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SKU	Supplier No.	Size	Your price	Quantity
BHC10900734	T0528	1x10^6 cells / 1.0 ml	-		Add to Quote

Immortalized Mouse Dendritic Cells (MutuDC1940)

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