Raji / Raji-luc Orthotopic Mouse Model Service for Lymphoma

The Raji / Raji-luc orthotopic mouse model for lymphoma engrafts an MHC class I-positive, Epstein-Barr virus-carrying human Burkitt lymphoma into the hematopoietic compartment of immunodeficient mice, where aggressive disseminated disease can be quantified non-invasively by bioluminescence. Focused exclusively on preclinical oncology, Alfa Cytology provides this platform as a complete service: authenticated Raji / Raji-luc cell supply, tail-vein orthotopic inoculation into SCID or NSG hosts, scheduled in vivo imaging and survival monitoring, and terminal bone marrow and organ-level analysis.

Overview of Raji / Raji-luc Orthotopic Mouse Model for Lymphoma

Raji holds a singular place in biomedical history: established in 1963 from a Burkitt lymphoma in an 11-year-old boy, it was the first continuous human hematopoietic cell line ever cultured, and it remains one of the most intensively used lines in existence. Registered with ATCC as CCL-86 and with DSMZ as ACC-319, Raji carries the defining Burkitt lesion—a chromosome 8;14 translocation generating a MYC-IGH fusion—and harbors Epstein-Barr virus in a latent, non-producer configuration: EBV nuclear antigen is expressed, but the genome carries deletions that prevent formation of infectious particles, so the line can be handled at biosafety level 1 even in genetic engineering work. Unlike its Burkitt cousin Daudi, Raji is beta-2-microglobulin positive and expresses surface MHC class I molecules, and it bears the EBV receptor CD21 while lacking surface immunoglobulin. Its karyotype, 46,XY with 4q⁺, 9p⁺, and 14q⁺ markers, shows unusual stability for a Burkitt line.

Fig 1. In vitro validation showing linear bioluminescence signal of Raji-luc cells exposed to luciferin (R² 0.997) and dose-dependent attenuation by isolated SCID/R2G2 mouse pelts, with the dark R2G2 dorsal pelt attenuating signal up to 120-fold—critical technical reference for quantitative BLI of Raji-luc disseminated lymphoma. (Hoegger, Mark J., et al., 2021)

In the orthotopic setting, Raji cells delivered intravenously engraft within the bone marrow of immunodeficient mice and generate an aggressive, leukemia-like disseminated disease—validation studies document whole-body bioluminescence accumulating in the pelvis, spine, ribs, skull, and spleen, with untreated cohorts reaching a roughly 20-day overall survival and terminating in progressive body-weight loss and hind-limb paralysis. Published inocula range from 1 × 10⁵ to 4.5 × 10⁶ luciferase-labeled cells, and Raji-luc has been the benchmark target in more than a decade of CD19 CAR-T and CAR-NK programs run in both SCID and NSG hosts, including humanized NSG-SGM3 systems for cytokine release syndrome research, CD37 CAR-T studies, dual CD19/CD20 constructs, and exosome-immune interaction work. The ATCC-engineered Raji-GFP-Luc2 reporter, verified to express high CD19 levels and to serve as a validated killing target for CD19 CAR-T cells, further standardizes the platform. Figure 1 outlines how this orthotopic lymphoma system is assembled and monitored.

Cell Line Information: Raji / Raji-luc

As the first continuously cultured human hematopoietic line, Raji has been characterized exhaustively across six decades. The defining attributes of the parent line and its luciferase-tagged derivatives are summarized below.

Our Services

A Raji / Raji-luc engagement at Alfa Cytology runs as one accountable pipeline: verified cell banking, standardized tail-vein orthotopic inoculation, calibrated IVIS imaging on the model’s compressed timeline, dosing of CAR-based or antibody-based therapeutics per the client’s regimen, and a terminal package combining survival statistics, ex vivo organ imaging, and bone marrow flow cytometry.

Workflow of Raji / Raji-luc Orthotopic Mouse Model Construction

Deploying the Raji / Raji-luc orthotopic lymphoma model is a rapid, highly reproducible workflow in which a small intravenous inoculum reliably generates disseminated bone marrow disease within days. The sequence below reflects the standard execution of this platform.

1. Cell Bank Verification and Expansion: Raji or Raji-luc stocks are authenticated by STR profiling and mycoplasma testing, then expanded in RPMI-1640-based medium with regular passage; clumping is minimized by gentle handling to ensure uniform single-cell suspensions.
2. Luciferase Calibration (Raji-luc Only): Signal output per cell is verified, with the ATCC Raji-GFP-Luc2 format providing a documented signal-to-noise ratio above 100 for high-confidence imaging.
3. Host Selection and Randomization: SCID, NSG, M-NSG, hIL-15 M-NSG, or humanized NSG-SGM3 mice are selected to match the therapeutic modality, acclimatized, and randomized by body weight under approved protocols.
4. Orthotopic Tail-Vein Inoculation: A single-cell suspension of luciferase-labeled Raji cells (published range 1 × 10⁵ to 4.5 × 10⁶ cells in a small sterile volume) is delivered via the lateral tail vein so that cells home to bone marrow and hematopoietic organs.
5. Engraftment Confirmation and Randomization by Radiance: Whole-animal bioluminescence after D-luciferin (150 mg/kg i.p.) confirms take within days; mice are then randomized by total body radiance to balance baseline tumor burden across groups.
6. Therapeutic Administration: CD19/CD20/CD37 CAR-T or CAR-NK cells, bispecific antibodies, or small molecules are dosed per the client’s schedule, typically within the first week after inoculation.
7. Longitudinal Imaging and Survival Endpoint: Twice-weekly IVIS sessions track dissemination, while body weight is recorded daily; hind-limb paralysis, ruffled fur, or 20% body-weight loss define the survival endpoint for Kaplan–Meier analysis.
8. Terminal Bone Marrow and Organ Analysis: At endpoint, bone marrow, spleen, liver, and blood are collected; tumor burden is quantified by flow cytometry for human B-cell markers, ex vivo bioluminescence, and H&E histopathology to complete the dataset.

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

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

Alfa Cytology has executed preclinical programs on the Raji / Raji-luc orthotopic platform for clients benchmarking CD19, CD20, and CD37 CAR constructs, evaluating bispecific antibodies and antibody-drug conjugates, and modeling cytokine release syndrome in humanized systems, delivering longitudinal bioluminescence records, survival analyses, and bone marrow flow-cytometry-supported endpoint packages. As each engagement is configured around the client’s own therapeutic concept, study outlines and representative datasets are available upon request.

Why Choose Alfa Cytology?

Raji / Raji-luc orthotopic studies move fast and tolerate no sloppiness: a poorly resuspended clumpy inoculum or a mistimed imaging session can blur group differences in a 20-day disease course. Alfa Cytology’s handling discipline and timeline calibration keep these studies crisp from first inoculation to final data package.

• Authenticated Raji / Raji-luc banks with STR and mycoplasma verification before in vivo use.
• Standardized tail-vein orthotopic inoculation with clump-controlled cell preparation for uniform engraftment.
• Radiance-based randomization balancing baseline tumor burden across treatment groups.
• Calibrated in-house IVIS imaging matched to the model’s compressed timeline.
• Flexible host selection (SCID, NSG, M-NSG, hIL-15 M-NSG, humanized NSG-SGM3) matched to each modality.
• Terminal bone marrow flow cytometry alongside ex vivo imaging and histopathology.

Contact Us

Whether the goal is to benchmark a CAR construct, evaluate a bispecific antibody, or model systemic B-cell malignancy, Alfa Cytology can stand up a Raji / Raji-luc orthotopic study on your timeline. Contact us today to scope your study design and dosing regimen, or reach out to our team for a customized quotation.

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