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Human Mast Cell Line (LAD2)

SKU : BHC10902116

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Applied Biological Materials (abm) Inc.

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Mfr.No.	T8157
Description	This novel mast cell line (LAD2) is derived from?D34+ cells obtained via marrow aspiration from a patient with aggressive mastocytosis, where no KIT mutations were identified. The cells are?haracterized by their slow proliferation, dependency on stem cell factor (SCF) for growth, and expression of functional surface Fc?RI receptors, which are crucial for studying mast cell biology. LAD2 cells closely resemble primary human mast cells in terms of growth characteristics, receptor expression, and functional responses, making them a valuable model for investigating allergic reactions, mast cell activation, and degranulation processes. Specifically, LAD2 has functional Fc?RI and FcyRI receptors and expresses CD4, 9, 13, 22, 45, 64, 71, 103, 117, 132, CCR5 and CXCR4, as well as CD14, 31 and 32, to a lesser degree. Cells are susceptible to T-tropic, M-tropic, and dual tropic HIV infection. LAD2 cells are widely used in research focused on allergy, immunology, and mast cell-related diseases. Their ability to degranulate in response to immunologic stimuli and their stable phenotype provide a reliable platform for testing the effects of various allergens, drugs, and therapeutic interventions. Additionally, LAD2 cells have been instrumental in studying the molecular mechanisms underlying mast cell activation and signaling pathways.Compared to LADR cells, LAD2 cells proliferate faster and are less granulated. LAD2 cells express a slightly different profile of surface markers and have lower tryptase expression compared to LADR cells, which influences their utility in specific research contexts.Note: understanding the recommended culture conditions and expected behaviour of this cell line are critical for establishing successful cultures.?dditional cell lines of interest:Cat. T8156 - Human Mast Cell Line (LADR)
Cell Type	Tumor Cells
Biosafety Level	2
Depositor	NIH (NIAID)
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	Male, 44, Mastocytosis
Expression Region	Fc?RI, CD33, CD34, CD63, CD117, CD133, CD193
Growth Properties	Suspension; Rounded, granulated; some large aggregates at higher densities
Growth Conditions	PriCoat?T25 Flasks (G299) are recommended for optimal cell culture. PriGrow X Series Medium for T8157 (TM8157) + 100ng/ml human Stem Cell Factor (Z100815) + 2mM L-Glutamine?G275) + 1% Penicillin/Streptomycin Solution (G255), 37.0?, 5% CO?.?eplace half of the medium with an equal volume of fresh medium, weekly. Do not allow cells to become over-confluent or grow past a density of 1x106?ells/ml.?ote: it is normal to observe cell death in the first two weeks after thawing. Allow cells to recover under the recommended culture conditons.?ells are slow-growing and extremely sensitive to mycoplasma contamination; routine testing should be performed.?Cells may lose functionality as a result of continuous culture.?egranulation assays must be conducted every 2 months to test functionality -protocol can be found under the "Documents" tab.
Tissue	Bone Marrow
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	Human (H. sapiens)
Cryopreservation	Following centrifugation, leave a small amount of culturing media (50-100 uL). Suspend cells in non-DMSO, non-FBS containing CryoScarless Cryopreservative (BioVerde) at a final concentration of 5x106?ells/mL. Place vials in a cryo container and store at -80? or in liquid nitrogen for longer preservation.

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 850rpm 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.6.?ost-thaw viability should be 70-95%; however, it is normal to observe increased cell death and debris in the first two-three weeks after the post-thaw period. Remove debris and dead cells during this period by skimming, where possible. Hemidepletion of the media should be performed 1-2 times per week depending on cell density. Allow cells to recover during the first 2-4 weeks post-thaw. In addition, add fresh 100 ng/ml SCF 1-2 times per week.
Subculture Protocol	1. Transfer expanding cultures to T75 or larger culture vessels to keep cell densities between 5x105??x105?ells/ml. Do not exceed a density of 1x106 cells/ml.?.?emove half the volume of the culture media and replace with fresh media (hemidepletion) the first or second week after thawing. Remove debris by skimming media, if possible. It is normal to see increased cell death for the first 2-3 weeks following thawing. 3. Alternatively, collect cells and centrifuge (850 rpm for 5 min ) to form a pellet. Discard half the supernatant and re-suspend the cell pellet in the remaining media. Add fresh complete media in 1:1 ratio and aliquot the cell suspension to new culture vessels, as desired. A complete media change is not advised.? 4. Incubate the cells at the recommended conditions.

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SKU	Supplier No.	Size	Your price	Quantity
BHC10902116	T8157	1x10^6 cells / 1.0 ml	-		Add to Quote

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