P388 / P388-luc Orthotopic Mouse Model Service for Leukemia

The P388 / P388-luc orthotopic mouse model seeds a classic murine lymphocytic leukemia into its natural systemic compartments of immunocompetent DBA/2 mice, reproducing the rapid multi-organ course of acute leukemia — peritoneal ascites, bone marrow infiltration, and splenomegaly — within days rather than weeks. Alfa Cytology supplies this proven platform as a complete preclinical service, from syngeneic host sourcing and intraperitoneal or intravenous inoculation to luciferase-based whole-body tracking in vivo and survival-based efficacy readouts, keeping your leukemia program moving at the pace its biology demands.

Overview of P388 / P388-luc Orthotopic Mouse Model for Leukemia

For hematologic malignancies, an orthotopic xenograft means transplanting tumor cells into the systemic compartment they normally occupy, and the P388 model is the archetype of this approach. P388 lymphocytic leukemia was induced in 1955 in a female DBA/2 mouse through skin painting with 3-methylcholanthrene, and together with L1210 it formed the foundation of the first large-scale cancer drug screening programs run by the National Cancer Institute — for decades, virtually every compound entering NCI evaluation was first tested against P388. The line retains an exceptionally aggressive phenotype: following inoculation of 10⁶ cells, tumor doubling times of 0.44 to 0.68 days and median animal survivals of 8 to 13 days are observed, depending on whether cells are given intraperitoneally, subcutaneously, intravenously, or intracerebrally. A family of drug-resistant derivatives — including P388/ADR (doxorubicin-selected), P388/DDP (cisplatin-selected), and multidrug-resistant P388/MDR — extends the model's usefulness to resistance and rescue-strategy research.

Fig 1. Longitudinal bioluminescence imaging of P388-derived P388D1.ffluc leukemia cells following intravenous inoculation, demonstrating systemic leukemia progression and prominent dissemination to the liver and spleen. (Lu, Lihui, et al., 2023)

In its classical configuration, P388 cells propagated in vitro are implanted intraperitoneally into syngeneic DBA/2 mice (or the hardier F1 hybrids B6D2F1 and CD2F1), where they first expand in the ascitic fluid and subsequently infiltrate the bone marrow, liver, and spleen; intravenous delivery instead seeds the bone marrow directly, mirroring the leukemic homing pattern seen in vivo. Because the hosts are immunocompetent, the model preserves tumor–immune interactions that human xenografts in immunodeficient mice cannot capture, making it valuable for immunotherapy, vaccine, and oncolytic strategies as well as conventional chemotherapy. The P388-luc variant, engineered for stable firefly luciferase expression after monoclonal selection, enables whole-body bioluminescence imaging in vivo that quantifies leukemia burden longitudinally, detects early organ engraftment, and sharpens the sensitivity of survival-based efficacy measurements — all within study timelines of roughly two to three weeks.

Cell Line Information: P388 / P388-luc

P388 (ATCC CCL-46) is one of the oldest and best-documented murine cancer cell lines. It originated in 1955 as a lymphocytic leukemia chemically induced by 3-methylcholanthrene in a female DBA/2 mouse, and it has been propagated ever since by intraperitoneal passage of diluted ascitic fluid containing 10⁶ cells per animal. In vitro, P388 grows in RPMI-1640-based media supplemented with 10% fetal bovine serum, L-glutamine, sodium pyruvate, and non-essential amino acids, showing adherent-type growth that contrasts with the suspension growth typical of most leukemia lines. Its benchmarked pharmacology — including well-characterized responses to standard agents and a rich panel of resistant sublines — makes P388 a reference standard against which new therapies can be compared. The P388-luc derivative carries a firefly luciferase reporter; as demonstrated for luciferase-tagged leukemia lines generally, monoclonal selection of high-signal clones is recommended to guarantee stable, comparable bioluminescent signals in vitro and in vivo.

Our Services

Speed and predictability are the twin currencies of leukemia research, and Alfa Cytology trades in both: P388 and P388-luc banks maintained under strict quality control, syngeneic host colonies ready on schedule, and imaging-calibrated randomization that places treatment arms on truly comparable disease burdens. One engagement covers inoculation, monitoring, dosing, and terminal organ analysis, so results arrive as a coherent package rather than a patchwork of subcontracted tasks.

Workflow of P388 / P388-luc Orthotopic Mouse Model Construction

The P388 orthotopic procedure is operationally simple but demands exacting cell preparation, because survival differences of a day or two translate into large efficacy effects at this growth rate. The workflow below describes construction and study execution from culture dish to final analysis.

1. Cell Expansion and Quality Control: P388 or P388-luc cells are expanded in vitro in RPMI-1640-based medium to the logarithmic phase; preparations are confirmed mycoplasma-negative and counted precisely, and luciferase expression is verified for tagged lines — with monoclonal, high-signal P388-luc clones preferred for stable bioluminescence in vitro and in vivo.
2. Host Selection and Acclimatization: Syngeneic DBA/2 mice, or the hardier B6D2F1/CD2F1 F1 hybrids for efficacy cohorts, typically 6–8 weeks of age, are acclimatized for approximately one week and health-screened before assignment to study groups.
3. Route Selection: The inoculation route is chosen to match the scientific question: intraperitoneal delivery recreates the classical ascitic leukemia, whereas intravenous (tail vein) delivery models disseminated disease with direct bone marrow homing; the standard challenge dose is 10⁶ viable cells per animal.
4. Inoculation: A single-cell suspension is prepared in sterile PBS and administered in a small volume (classically 0.1 mL intraperitoneally, or ~0.2 mL intravenously) with verified cell concentration, recording implantation day as day 0.
5. Disease Monitoring: Animals are checked daily for body weight, posture, and activity; for P388-luc studies, whole-body bioluminescence imaging in vivo is performed twice weekly after intraperitoneal D-luciferin (150 mg/kg), with peripheral blood smears prepared as disease advances.
6. Randomization and Treatment: Mice are assigned to vehicle and treatment arms at a defined time point or bioluminescence threshold, ensuring matched starting burdens, and test articles are dosed on the client-specified schedule with toxicology-relevant observations recorded throughout.
7. Survival Follow-Up and Endpoint Analysis: Animals are followed to a humane moribund endpoint or a predefined study day; at termination, spleen and liver are weighed, bone marrow is flushed from the femurs, and blood is collected for smear and flow cytometric analysis, with ex vivo organ imaging confirming signal distribution in P388-luc studies.

Fig 2. P388 / P388-luc orthotopic mouse model construction workflow.

Case Study-P388 / P388-luc Orthotopic Mouse Model Development

The P388 system has underpinned preclinical leukemia research for generations, from classic single-agent chemotherapy evaluations to modern applications such as multidrug-resistance reversal, oncolytic therapy testing, and immune-based approaches in immunocompetent hosts. Alfa Cytology has conducted client studies on this platform spanning cytotoxic agents, resistance-modulating combinations, and biologic therapeutics, with readouts combining survival analysis, longitudinal bioluminescence imaging in vivo, and terminal flow cytometric assessment of marrow and spleen involvement. Anonymized study summaries and example data formats are available on request.

Why Choose Alfa Cytology?

Few models carry the validation depth of P388, but realizing that depth in practice requires disciplined execution at a pace most CROs rarely work at. Alfa Cytology has built its leukemia service around exactly that discipline.

• A model with seven decades of published benchmark data behind every efficacy measurement you will generate.
• Flexible inoculation routes — intraperitoneal ascitic disease or intravenous bone-marrow-seeding dissemination — selected to fit your mechanism of interest.
• Authenticated P388 banks plus resistant counterparts (P388/ADR, P388/DDP, P388/MDR) for combination and rescue-strategy studies.
• Quality-controlled P388-luc with monoclonal luciferase selection for stable, comparable bioluminescence in vivo.
• Immunocompetent syngeneic hosts that preserve tumor–immune interactions unavailable in human xenograft systems.
• Compact study timelines of roughly two to three weeks from inoculation to survival curves and organ-level analyses.
• Integrated flow cytometry, histopathology, and ex vivo imaging endpoints from a single study team.

Contact Us

If your leukemia program needs a fast, rigorously benchmarked in vivo system, the P388 / P388-luc orthotopic model is a proven place to start — and Alfa Cytology is ready to run it for you. Contact us to discuss inoculum design, resistant subline options, or imaging-enabled study formats, and receive a tailored quotation from our leukemia modeling team.

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