| Field | Specification |
|---|---|
| Product Format | Frozen |
| Product Type | |
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Overview
The LAD2 human mast cell line originates from bone marrow aspirates of a patient with mast cell sarcoma/leukemia. These cells exhibit ultrastructural characteristics of human mast cells and express FcεRI, along with markers such as CD4, 9, 13, 14, 22, 31, 32, 45, 64, 71, 103, 117, 132, CXCR4 (CD184), and CCR5 (CD195).
Key elements and design rationale
- Model identity: Human Mast Cell Line (LAD2) ; Capable of histamine release (degranulation) is supplied as a tumor cell line derived from Human bone marrow.
- Growth properties: Suspension; Rounded, granulated; some large aggregates at higher densities
- Growth conditions: Note: understanding the recommended culture conditions and expected behaviour of this cell line are critical for establishing successful cultures. PriGrow X Series Medium for T8157 (TM8157) + 100 ng/ml human Stem Cell Factor (Z100815) + 2 mM L-glutamine (G275) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. Replace 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 cells/ml. Note: it is normal to observe cell death in the first two weeks after thawing. Allow cells to recover under the recommended culture conditons. Cells are slow-growing and extremely sensitive to mycoplasma contamination; routine testing should be performed. *Cells may lose functionality as a result of continuous culture. Degranulation assays must be conducted every 2 months to test functionality. For optimal degranulation, 1 million LAD2 and LADR cells should be cultured in hSCF-free media for 4 hours prior to sensitization. The cells should then be sensitized with human IgE (Y70000), followed by IgE crosslinking (Y79000) to stimulate degranulation before performing the Mast Granulation Assay (G7800). For more details refer to Mast Cell Preparation for Granulation Assay - Guideline.
- Product format: Frozen, BSL-2
This cell-based model is generally used in immunology, hematology, and signaling studies. Donor/background information is available for contextual interpretation.
Biological background
Additionally, they contain intracytoplasmic histamine, tryptase, and chymase. Notably, LAD2 cells do not carry activating mutations at codon 816 of the c-kit gene. Similar to LAD1, LAD2 cells release β-hexosaminidase upon FcεRI or FcϒRI aggregation.The availability of the LAD2 mast cell line provides a unique opportunity to study the biology of human mast cells. Over time, it has become the most widely used and recognized gold-standard mast cell line for allergy and mastocytosis research, as well as a crucial tool in pharmaceutical development. Donor/background information provided for this product: Male, 44, Mastocytosis.
Research relevance and current trends
- Cell-line models continue to be used for tumor phenotype comparison, pathway perturbation studies, and assay development in controlled in vitro systems.
- Engineered and subtype-defined tumor lines are often used to compare growth behavior, reporter output, and response patterns across matched experimental conditions.
- When metastatic or lineage features are described, investigators commonly interpret results alongside morphology, passage history, and culture environment.
Common research applications
- Cancer biology studies that compare proliferation-associated behavior, morphology, and pathway responses in vitro.
- Assay development for treatment response, reporter monitoring, or phenotype comparison under matched culture conditions.
- Side-by-side comparison of engineered versus parental background characteristics when relevant to the study design.
Changes in morphology, growth rate, viability, or reporter signal are typically interpreted together with passage history, culture matrix, and the specified growth conditions for the model.
Notes for experimental interpretation
- Morphology, doubling behavior, and reporter or marker output can shift with passage number, substrate choice, and medium composition; these variables should be recorded alongside experimental readouts.
- Matched controls such as parental cells, untreated cultures, or parallel cultures maintained under identical conditions help distinguish background effects from biology of interest.
Culture and product details
- Growth Conditions: Note: understanding the recommended culture conditions and expected behaviour of this cell line are critical for establishing successful cultures. PriGrow X Series Medium for T8157 (TM8157) + 100 ng/ml human Stem Cell Factor (Z100815) + 2 mM L-glutamine (G275) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. Replace 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 cells/ml. Note: it is normal to observe cell death in the first two weeks after thawing. Allow cells to recover under the recommended culture conditons. Cells are slow-growing and extremely sensitive to mycoplasma contamination; routine testing should be performed. *Cells may lose functionality as a result of continuous culture. Degranulation assays must be conducted every 2 months to test functionality. For optimal degranulation, 1 million LAD2 and LADR cells should be cultured in hSCF-free media for 4 hours prior to sensitization. The cells should then be sensitized with human IgE (Y70000), followed by IgE crosslinking (Y79000) to stimulate degranulation before performing the Mast Granulation Assay (G7800). For more details refer to Mast Cell Preparation for Granulation Assay - Guideline.
- Seeding Density (cells/ml): 500,000-800,000
- Thaw cells quickly in a 37°C water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination.
- 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.
- 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.
- 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.
- Incubate the cells at the recommended conditions.
- Post-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. **It’s normal for some cells to adhere to the vessel walls. Gently tapping the culture vessel should help dislodge and resuspend them for continued culturing.** †Skimming refers to the method of removing floating debris and dead cells from the culture media. In cell culture, "skimming" might involve gently removing the upper layer of the media where dead cells and debris tend to accumulate, typically using a pipette or similar instrument. This helps maintain the quality of the culture environment, promoting better recovery and growth of viable cells.‡Hemidepletion of media refers to the partial removal of media to refresh the culture environment, control cell density, and promote healthy cell growth. This process is important for maintaining a successful culture of the mast cell line.
- Transfer expanding cultures to T75 or larger culture vessels to keep cell densities between 5x105 - 8x105 cells/ml. Do not exceed a density of 1x106 cells/ml.
- Remove 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.
- 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.
- Incubate the cells at the recommended conditions.
How should I handle live cells once I receive them?
https://www.abmgood.com/immortalized-cells-documents.html
Following these guidelines will help ensure optimal cell viability and performance.
Why are these cells classified as biosafety level II?
What is your warranty or return policy?
Please refer to the following link for full information:
https://www.abmgood.com/terms
For additional questions, our Order team is happy to assist and can be reached at order@abmgood.com.
How many times can cells divide?
Primary cells have a limited lifespan and will undergo a finite number of population doublings before entering senescence. The exact number varies by cell type and culture conditions.
Immortalized cell lines are capable of extended or indefinite proliferation under proper culture conditions, although growth characteristics may vary between lines.
Do I need Applied Cell Extracellular Matrix (G422) if I am using PriCoat™ flasks?
Cell line sourcing and selection (species, tissue, and disease model matching) · Stable cell line engineering (overexpression, knockdown, knockout via CRISPR/Cas9, shRNA, sgRNA) · Reporter gene integration (GFP, RFP, luciferase, fluorescent/bioluminescent constructs) · Genome editing and knockin (point mutations, tagged endogenous proteins, conditional alleles) · Inducible expression systems (Tet-On/Off and regulatable constructs) · Drug resistance marker selection (puromycin, G418, hygromycin, and others) · Custom growth and media optimisation for specific assay requirements · Scale-up production for high-throughput screening campaigns · Authentication and QC services (STR profiling, mycoplasma testing, viability assessment). Talk to a Scientist or contact support@biohippo.com.
Kirshenbaum, A. S., Akin, C., Wu, Y., Rottem, M., Goff, J. P., Beaven, M. A., ... & Metcalfe, D. D. (2003). Characterization of novel stem cell factor responsive human mast cell lines LAD 1 and 2 established from a patient with mast cell sarcoma/leukemia; activation following aggregation of FcεRI or FcγRI. Leukemia research, 27(8), 677-682.
Kirshenbaum, A. S., Yin, Y., Sundstrom, J. B., Bandara, G., & Metcalfe, D. D. (2019). Description and Characterization of a Novel Human Mast Cell Line for Scientific Study. International journal of molecular sciences, 20(22), 5520. https://doi.org/10.3390/ijms20225520
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