MOLM-13 / MOLM-13-luc Orthotopic Mouse Model Service for Leukemia

The MOLM-13 / MOLM-13-luc orthotopic mouse model of leukemia reproduces an MLL-rearranged, FLT3-mutated human acute myeloid leukemia in its native hematopoietic niche, with luciferase bioluminescence converting systemic disease burden into a quantitative, serially measurable signal. Alfa Cytology provides this platform as a full preclinical service — covering authenticated MOLM-13 banking, luciferase line generation and validation, tail-vein engraftment in NSG or NCG recipients, scheduled in vivo imaging, survival scoring, and flow-cytometric human engraftment analysis — so sponsors can interrogate FLT3-pathway, epigenetic, and cell-therapy strategies with time-resolved in vivo data.

Overview of MOLM-13 / MOLM-13-luc Orthotopic Mouse Model for Leukemia

MOLM-13 is a human acute myeloid leukemia cell line established in 1995 from the peripheral blood of a 20-year-old male patient whose AML FAB M5a disease — acute monocytic leukemia — had relapsed after an initial myelodysplastic syndrome of the refractory-anemia-with-excess-blasts type. The line is genetically defined by a KMT2A-MLLT3 (MLL-AF9) fusion arising from a cryptic insertion rather than a simple translocation, an internal tandem duplication in FLT3, and a CBL deltaExon8 mutation; its immunophenotype (CD34+, CD33+, CD15+, CD4+, cytoplasmic CD68+, with HLA-DR and CD13 negative) mirrors the monocytic blast population of the original patient. These lesions sit squarely within the molecular landscape of adverse-risk human AML, which is why MOLM-13 has become a preferred in vitro and in vivo reference for studying MLL-fusion biology, FLT3 signaling dependence, and differentiation-blockade reversal.

Fig 1. Longitudinal bioluminescence monitoring of an orthotopic MOLM-13-Luc AML xenograft established by intravenous engraftment, demonstrating dose-dependent leukemia regression following pharmacological c-Myc degradation. (Xu, Ying, et al., 2022)

The orthotopic application of this line follows the clinical route of leukemic dissemination: labeled cells are delivered intravenously into severely immunodeficient recipients, home preferentially to the bone marrow, and then expand through blood, spleen, and liver until normal hematopoiesis is overwhelmed. Firefly luciferase-tagged derivatives — constructed by retroviral or lentiviral transfer, in some configurations co-expressing GFP or mCherry — make it possible to quantify whole-body leukemia burden by bioluminescence imaging after D-luciferin administration, with photon flux dynamics faithfully reflecting engraftment, expansion, and treatment response. This imaging window has powered a broad portfolio of preclinical studies on the MOLM-13-luc platform, including FLT3 siRNA nanoparticle delivery, c-Myc and ribogenesis inhibitors, epigenetic modulators, CAR-NK cell therapy, and donor-derived Vγ9Vδ2 T-cell approaches, all monitored longitudinally in vivo.

Cell Line Information: MOLM-13 / MOLM-13-luc

MOLM-13 is a molecularly well-annotated human AML line, and its luciferase-tagged derivative extends it into the optical imaging domain. Core attributes are listed below.

Our Services

Sponsors turn to Alfa Cytology for MOLM-13-luc work because every detail that influences an aggressive AML readout is controlled in-house: authenticated ACC 554 stocks handled at low passage, reporter lines titrated for uniform luciferase output before release into studies, and imaging calendars synchronized to the engraftment curve so treatment windows are never missed. Beyond operations, Alfa Cytology contributes scientific steering — advising on challenge dose, randomization thresholds, and endpoint panels (including human CD45/CD33 engraftment analysis) — so that each MOLM-13-luc orthotopic study lands cleanly on its efficacy question.

Workflow of MOLM-13 / MOLM-13-luc Orthotopic Mouse Model Construction

The MOLM-13-luc orthotopic study is engineered around a short, steep disease curve: engraftment is fast, burden doubles quickly, and treatment windows open and close within days. The workflow below keeps every phase — cell release, intravenous delivery, imaging, and analysis — on a tightly scheduled track.

1. Line release and reporter validation: MOLM-13 / MOLM-13-luc vials are expanded from authenticated master banks, cleared of mycoplasma, and released into studies only after confirming homogeneous luciferase expression by in vitro luminescence titration across serial passages.
2. Engraftment cell preparation: Study cultures maintained at 0.4-2.0×10⁶ cells/mL are harvested in log phase, washed, counted, and resuspended in serum-free medium at the challenge dose (commonly 2×10⁵ to 5×10⁵ cells in approximately 100 µL per mouse).
3. Recipient preparation and tail-vein inoculation: NSG or NCG mice are acclimatized, stratified by body weight, and injected intravenously via the lateral tail vein using warming and transillumination to secure vessel access and consistent delivery.
4. Engraftment verification and baseline imaging: Several days after inoculation, mice receive intraperitoneal D-luciferin (typically 150-300 mg/kg) and whole-body bioluminescence imaging verifies bone-marrow homing and quantifies baseline leukemia burden.
5. Signal-based randomization and treatment start: Animals are allocated to arms with matched photon flux, and dosing commences on schedule — small molecules by oral gavage or intraperitoneal injection, nucleic-acid complexes intraperitoneally, or cell therapies (e.g., CAR-NK or Vγ9Vδ2 T cells) intravenously.
6. Serial burden monitoring: Whole-body imaging is repeated at protocol-defined intervals, with body weight and clinical condition scored in parallel, generating per-animal disease-progression curves and early evidence of response or relapse.
7. Human engraftment analysis and survival endpoints: Peripheral blood, bone marrow, and spleen are sampled for human CD45/CD33 flow cytometry to quantify leukemic chimerism; morbidity-defined survival is recorded for Kaplan-Meier analysis, with spleen weights and histopathology documenting terminal disease.

Fig 2. MOLM-13 / MOLM-13-luc orthotopic mouse model construction workflow.

Case Study-MOLM-13 / MOLM-13-luc Orthotopic Mouse Model Development

Alfa Cytology has implemented the MOLM-13-luc orthotopic platform across a range of sponsor programs spanning targeted small molecules, nucleic-acid delivery systems, and adoptive cell therapies. In these studies, serial whole-body bioluminescence captured the steep expansion kinetics of engrafted leukemia and quantified treatment-induced suppression of systemic burden, while terminal human CD45/CD33 flow-cytometric analysis confirmed engraftment levels across blood, bone marrow, and spleen and survival data were resolved with Kaplan-Meier methods. Where programs required it, bone-marrow infiltration was further assessed by femoral sampling and luminescence readout of marrow cell suspensions. Program-specific parameters and outcomes are held as sponsor-specific deliverables, and representative outlines can be provided on request.

Why Choose Alfa Cytology?

MOLM-13-luc studies fail quietly when cell quality, timing, or imaging discipline slips — and succeed when all three are managed deliberately. Alfa Cytology’s service is organized around that principle, giving sponsors a dependable, fast-turnaround AML platform. Highlights include:

• Authenticated DSMZ ACC 554 stocks expanded at low passage with verified reporter output before study release.
• Severely immunodeficient NSG/NCG hosts enabling high-penetrance human leukemia engraftment.
• Signal-matched randomization that neutralizes engraftment variability across aggressive disease courses.
• Imaging cadences calibrated to MOLM-13 growth kinetics so treatment effects are captured, not missed.
• Human CD45/CD33 flow-cytometric engraftment analysis integrated as a standard endpoint.
• Support for diverse modalities, from oral and intraperitoneal small molecules to CAR-NK and Vγ9Vδ2 T-cell dosing.
• A dedicated study manager providing timepoint-by-timepoint visibility from inoculation to final report.

Contact Us

Seeking an imaging-ready human AML model for your next preclinical milestone? Contact us to walk through challenge doses, schedules, and endpoint options for the MOLM-13 / MOLM-13-luc platform. You can also reach out to Alfa Cytology directly for a tailored proposal and quotation — our leukemia team will respond with a study design built to your timeline.

For research use only. Not intended for any clinical use.