OCI-AML3 Xenograft Rat Model Service for Leukemia

The OCI-AML3 xenograft rat model captures the defining molecular signature of NPM1-mutated acute myeloid leukemia—complete with aberrant cytoplasmic nucleophosmin expression and DNMT3A R882C co-mutation—within an immunodeficient rat host that affords expanded blood volumes for pharmacokinetic profiling and deep tissue sampling for biomarker discovery. Alfa Cytology constructs this model using authenticated DSMZ ACC 582 cell stocks and validated nude rat engraftment protocols, delivering reproducible leukemic burden kinetics and integrated pharmacodynamic endpoints that advance your preclinical AML therapeutic pipeline with molecular precision.

Overview of OCI-AML3 Xenograft Rat Model for Leukemia

The OCI-AML3 cell line (DSMZ ACC 582) was established in 1987 from the peripheral blood of a 57-year-old man diagnosed with acute myeloid leukemia of the French-American-British (FAB) M4 subtype (acute myelomonocytic leukemia). Genetically, the line carries the hallmark NPM1 exon-12 insertion mutation (Type A, TCTG duplication), which drives aberrant cytoplasmic accumulation of nucleophosmin protein—an immunocytochemical hallmark observed in approximately 35% of adult AML cases and associated with normal karyotype, CD34 negativity, and favorable response to induction chemotherapy. In addition, OCI-AML3 harbors the DNMT3A R882C dominant-negative mutation and an NRAS Q61L activating mutation, together recapitulating a common co-mutational landscape seen in primary NPM1-mutated AML. The cells exhibit a myelomonocytic immunophenotype (CD13+, CD14+, CD15+, CD33+, CD34+) and a doubling time of approximately 30–40 hours under standard culture conditions.

Fig 1. OCI-AML3 Luc/GFP xenograft in vivo imaging (Panobinostat, Adavosertib), mtNPM1 PDX survival curve. (Mill, Christopher P., et al., 2023)

In vivo, OCI-AML3 readily engrafts into immunodeficient hosts, maintaining its characteristic NPM1 cytoplasmic expression and myelomonocytic differentiation markers throughout serial passage. When implanted into nude rats, the model leverages the larger physiological scale of the host to enable repeated peripheral blood sampling for pharmacokinetic analysis, expanded bone marrow and spleen harvest for flow cytometric blast quantification, and sufficient tumor tissue for multiplex biomarker and epigenetic profiling. The model has been extensively utilized to evaluate CXCR4 antagonists that disrupt leukemia–stromal niche interactions, KIT inhibitors targeting SCF-dependent survival signaling, and combination regimens pairing hypomethylating agents with targeted therapeutics. Its faithful retention of the NPM1-mutant phenotype makes OCI-AML3 an indispensable preclinical platform for adult AML drug development.

Cell Line Information: OCI-AML3

The table below summarizes the authenticated characteristics of the OCI-AML3 human acute myeloid leukemia cell line, compiled from DSMZ repository data, Cellosaurus records, and peer-reviewed literature.

Our Services

Alfa Cytology bridges the gap between in vitro OCI-AML3 characterization and in vivo translational outcomes by managing every stage of xenograft construction—from authenticated cell banking and mycoplasma-free expansion through intravenous or subcutaneous implantation into immunodeficient rats, longitudinal disease burden monitoring via bioluminescence and flow cytometry, serial blood collection for pharmacokinetic analysis, and terminal histopathology with quantitative biomarker readouts. Each study is tailored to your compound's mechanism of action and conducted under accredited IACUC oversight with full GLP-aligned documentation.

Workflow of OCI-AML3 Xenograft Rat Model Construction

Construction of the OCI-AML3 xenograft rat model follows a standardized yet adaptable workflow designed to achieve high engraftment rates, consistent leukemic burden kinetics, and robust pharmacodynamic endpoints. The protocol supports both subcutaneous flank implantation for localized tumor mass monitoring and intravenous delivery for disseminated leukemia modeling, selected according to the scientific objective.

1. Cell Line Resuscitation & Quality Control: Cryopreserved OCI-AML3 stocks (DSMZ ACC 582) are thawed and expanded in alpha-MEM supplemented with 10–20% heat-inactivated FBS under antibiotic-free, low-passage conditions. Cell identity is confirmed by morphology, growth curve analysis, and STR profiling against the authenticated reference (RRID: CVCL_1844). Mycoplasma testing is performed prior to in vivo use. Harvest occurs during exponential phase (0.5–2.0 × 10⁶ cells/mL); viability is assessed by trypan blue exclusion, with only suspensions exceeding 98% viability advanced to implantation. Cells are washed twice in PBS and resuspended at 5 × 10⁶ cells per 100 µL for subcutaneous injection, or at 2 × 10⁶ cells per 200 µL for intravenous tail-vein delivery.
2. Host Selection & Conditioning: Immunodeficient nude rats (Crl:NIH-Foxn1rnu, female, 6–8 weeks old, 150–200 g) are ordered from accredited vendors and acclimated for 7–10 days under specific-pathogen-free conditions. Baseline body weights and complete blood counts are recorded. Where disseminated leukemia modeling is required, rats may receive sublethal conditioning (low-dose busulfan or total body irradiation at 2.25 Gy) 24 hours prior to cell injection to enhance bone marrow engraftment, following established AML xenotransplantation protocols.
3. Leukemic Cell Implantation: For subcutaneous models, 100 µL of the OCI-AML3 single-cell suspension (5 × 10⁶ cells) is injected into the right flank using a 25-gauge needle. For systemic leukemia models, 200 µL of cell suspension (2 × 10⁶ cells) is delivered via tail-vein injection. The subcutaneous route generates a localized tumor mass amenable to caliper measurement and intratumoral dosing, while the intravenous route models hematogenous dissemination with subsequent bone marrow, spleen, and liver infiltration.
4. Post-Implantation Monitoring & Disease Tracking: Animals are monitored daily for clinical signs of leukemia progression including ruffled coat, hunched posture, reduced motility, and hind-limb paralysis in disseminated models. Body weights are recorded three times weekly. Subcutaneous tumors are measured twice weekly with digital calipers (volume = L × W² / 2) from first palpable detection, typically 7–10 days post-implantation. For disseminated models, peripheral blood is collected via retro-orbital bleeding or saphenous vein puncture for human CD45+ blast quantification by flow cytometry. Optional bioluminescence imaging is employed for luciferase-transduced OCI-AML3 variants.
5. Therapeutic Intervention & Endpoint Harvest: Upon reaching the target tumor volume (typically 100–200 mm³ for subcutaneous) or defined disease burden in disseminated models, animals are randomized into treatment and vehicle control cohorts. Test articles are administered via the sponsor-specified route—intravenous, oral gavage, intraperitoneal, or intratumoral—according to the dosing regimen. Tumor measurements and body weights continue on schedule until the study endpoint. At termination, animals are humanely euthanized. Subcutaneous tumors, bone marrow, spleen, liver, lungs, and blood are harvested. Tumor weight is recorded, tissues are digitally imaged, and samples are processed for H&E histopathology, Ki-67 proliferation index, TUNEL apoptosis scoring, NPM1 cytoplasmic expression verification by immunohistochemistry, and plasma drug-concentration profiling.

Fig 2. OCI-AML3 Xenograft Rat Model construction workflow.

Case Study-OCI-AML3 Xenograft Rat Model Development

In a recent preclinical engagement, Alfa Cytology utilized the OCI-AML3 disseminated xenograft rat model to evaluate the antileukemia efficacy of a novel CXCR4 antagonist candidate designed to disrupt leukemia–bone marrow stromal niche interactions. Following authenticated OCI-AML3 cell expansion and intravenous injection into conditioned nude rats, leukemic engraftment was confirmed by peripheral blood flow cytometry for human CD45+ blasts prior to randomization into vehicle, benchmark, and escalating dose cohorts. The candidate compound was administered via daily subcutaneous injection for 21 days, with longitudinal disease burden monitoring through serial bioluminescence imaging and interim flow cytometric blast quantification. Terminal analyses included bone marrow and spleen immunohistochemistry for human CD45 and NPM1 cytoplasmic expression, Ki-67 and TUNEL dual staining to assess proliferation and apoptosis, and plasma pharmacokinetic profiling. The dataset revealed dose-dependent mobilization of leukemic cells from the bone marrow niche accompanied by reduced tissue infiltration and prolonged survival signals, providing the sponsor with pharmacodynamic evidence to support downstream candidate optimization and IND-enabling toxicology planning.

Why Choose Alfa Cytology?

Partnering with Alfa Cytology for your OCI-AML3 acute myeloid leukemia program means accessing a specialized preclinical infrastructure built around authenticated leukemia cell biology, immunodeficient rat surgical expertise, and integrated pharmacodynamic analytics tailored to NPM1-mutant disease.

• Authenticated OCI-AML3 master and working cell banks are maintained under documented low-passage protocols with periodic STR verification and mycoplasma screening to ensure NPM1 and DNMT3A mutational stability and consistent engraftment.
• Our surgical team is proficient in both subcutaneous flank and tail-vein intravenous implantation techniques in immunodeficient rats, achieving high engraftment rates with minimal procedure-related morbidity.
• The larger physiological scale of rats enables serial blood draws for pharmacokinetic profiling, expanded bone marrow and spleen harvest for flow cytometric blast quantification, and sufficient tissue for multiplex biomarker and epigenetic analysis.
• Integrated molecular and histopathology capabilities include NPM1 cytoplasmic expression verification, DNMT3A activity assays, human CD45+ engraftment quantification by flow cytometry, and Ki-67/TUNEL scoring.
• Study protocols are customized to your therapeutic modality—whether CXCR4 antagonists, KIT inhibitors, hypomethylating agents, or combination regimens—with clear pharmacodynamic and efficacy decision criteria.
• All in vivo work is conducted under fully accredited IACUC oversight with real-time veterinary monitoring and GLP-aligned documentation, ensuring ethical integrity and generating audit-ready data packages for regulatory submissions.

Contact Us

If your acute myeloid leukemia therapeutic pipeline demands a preclinical model that faithfully recapitulates NPM1-mutated, DNMT3A-co-mutated disease biology within an immunodeficient rat host, reach out to us to discuss how Alfa Cytology can architect an OCI-AML3 study tailored to your development milestones. Our scientific team will review your target profile, propose a customized protocol with integrated pharmacokinetic and pharmacodynamic endpoints, and deliver a comprehensive proposal within two business days. Contact us today and accelerate your preclinical proof-of-concept with a CRO that understands the complexities of NPM1-mutated leukemia modeling.

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