5TGM1 / 5TGM1-luc Orthotopic Mouse Model Service for Multiple Myeloma (MM)

The 5TGM1 / 5TGM1-luc orthotopic mouse model for multiple myeloma recreates bone marrow-homing plasma cell disease with clinically relevant osteolytic bone lesions in syngeneic immunocompetent hosts, with luciferase labeling enabling non-invasive quantification of tumor burden. Operating exclusively in preclinical oncology, Alfa Cytology delivers this myeloma platform end to end: authenticated 5TGM1 / 5TGM1-luc cell supply, tail-vein orthotopic inoculation into C57BL/KaLwRij or NSG recipients, scheduled bioluminescence imaging paired with serum IgG2b monitoring, and terminal bone microCT/DEXA analysis.

Overview of 5TGM1 / 5TGM1-luc Orthotopic Mouse Model for Multiple Myeloma (MM)

The 5TGM1 line descends from one of the most instructive model systems in hematologic malignancy research. In the late 1970s, Radl and colleagues observed that aged C57BL/KaLwRij mice spontaneously develop a monoclonal gammopathy resembling human MGUS that can progress to frank myeloma, and they established the 5T series by serial transplantation of bone marrow from these animals. From the aggressive 5T33 subline, Garrett and colleagues generated 5TGM1 in 1997 through continuous in vivo passage, creating a cell line that grows reliably in vitro and retains its marrow-homing identity. The cells are CD138⁺ plasma cells that secrete a monoclonal IgG2b paraprotein—providing a serum biomarker that mirrors the M-spike of human disease—and carry Trp53 and Kdm6a mutations with aneuploid gains of chromosomes 15 and 18 and frequent loss of chromosome 16, a mutational spectrum with genuine overlap with human myeloma.

Fig 1. Comprehensive 5TGM1-Luc model study combining in vivo bioluminescence imaging (dorsal/ventral views, weeks 1–5) with serum IgG2b quantification, bone micro-CT, and IgG2b immunohistochemistry of femurs, showing that the MMP-2 inhibitor ML104 (but not ML111/ML115) reduced 5TGM1-Luc tumor burden and osteolytic lesions compared with vehicle and zoledronic acid (ZOL). (Shay, Gemma, et al., 2017)

Orthotopically, 5TGM1 delivered intravenously homes to bone marrow and spleen, producing disseminated disease with pronounced osteolytic lesions in vertebrae and long bones—recapitulating myeloma bone disease, the dominant clinical complication of the human disease. The standard protocol injects 1 × 10⁶ cells into C57BL/KaLwRij mice, where engraftment is typically confirmed by bioluminescence around week four and untreated animals reach endpoints within 3–5 weeks with documented median survival near 44.5 days; in NSG hosts the same inoculum engrafts earlier (week two) and animals survive longer (median ~56 days) while developing more severe bone loss. Luciferase-tagged derivatives (5TGM1-luc) enable weekly whole-body bioluminescence after D-luciferin administration, with ex vivo imaging confirming signal across long bones, spine, and spleen; GFP formats such as 5TGM1-eGFP have maintained stable expression for over a year in continuous culture. The model has validated immunotherapies—anti-PD-1 and anti-CD137 are active in the syngeneic setting, whereas proteasome inhibitor responses differ between nude and syngeneic hosts, underscoring the value of the immune-intact format—and has served radiopharmaceutical, bisphosphonate, and dormancy-niche programs.

Cell Line Information: 5TGM1 / 5TGM1-luc

5TGM1 has been refined across decades of transplantable myeloma research, and its luciferase-tagged derivatives have become the default imaging standard for syngeneic myeloma studies. The defining attributes are summarized below.

Our Services

A 5TGM1 / 5TGM1-luc engagement with Alfa Cytology is run as one continuous program: verified cell banking, tail-vein orthotopic inoculation into syngeneic or immunodeficient recipients, weekly IVIS imaging paired with serum IgG2b curves, dosing per the client’s regimen, and a terminal package combining microCT/DEXA bone analysis, ex vivo organ imaging, and bone marrow flow cytometry.

Workflow of 5TGM1 / 5TGM1-luc Orthotopic Mouse Model Construction

Building a 5TGM1 or 5TGM1-luc orthotopic myeloma model is a bone-focused workflow in which intravenous inoculation seeds marrow, imaging quantifies burden, and terminal microCT quantifies the osteolytic damage that defines the disease. The sequence below reflects the standard execution of this platform.

1. Cell Bank Verification and Expansion: 5TGM1 or 5TGM1-luc stocks are authenticated, mycoplasma-tested, and expanded in RPMI-1640-based medium, maintained at 0.5–2 × 10⁶ cells/mL with viability confirmed by trypan exclusion.
2. Luciferase Calibration (5TGM1-luc Only): Signal output per cell is verified in vitro, and CD138⁺ identity is confirmed by flow cytometry before in vivo use.
3. Host Preparation and Randomization: Eight- to twelve-week-old C57BL/KaLwRij (syngeneic) or NSG mice are acclimatized and randomized by body weight under approved protocols.
4. Orthotopic Tail-Vein Inoculation: 1 × 10⁶ cells (greater than 90% viability) in 100 µl of sterile PBS are delivered via the tail vein so that plasma cells home to bone marrow and spleen.
5. Engraftment Confirmation and Longitudinal Imaging: Weekly whole-animal bioluminescence after intraperitoneal D-luciferin (1.5 mg/mouse, 10-minute delay) tracks marrow and splenic burden from as early as week two in NSG or week four in syngeneic hosts; concurrent serum samples are collected for IgG2b ELISA.
6. Treatment Administration: Test agents—proteasome inhibitors, bisphosphonates, immunotherapies, or radiopharmaceuticals—are dosed per the client’s schedule once engraftment is confirmed.
7. Surveillance and Survival Endpoint: Mice are monitored daily; hind-limb paralysis, significant activity reduction, or 15–20% body-weight loss define the survival endpoint for Kaplan–Meier analysis.
8. Terminal Bone and Organ Analysis: At endpoint, spines and femur/tibia are scanned ex vivo by microCT (BV/TV, trabecular thickness) and DEXA; bone marrow, spleen, and other organs are imaged ex vivo and profiled by flow cytometry, with histopathology confirming osteolytic lesions.

Fig 2. 5TGM1 / 5TGM1-luc orthotopic mouse model construction workflow.

Case Study-5TGM1 / 5TGM1-luc Orthotopic Mouse Model Development

Alfa Cytology has executed preclinical programs on the 5TGM1 / 5TGM1-luc orthotopic platform for clients evaluating proteasome inhibitors, immunotherapies, bisphosphonates, and bone-targeted radiopharmaceuticals, delivering weekly bioluminescence growth curves, serum IgG2b biomarker data, survival analyses, and microCT-supported bone disease quantification. Because each engagement is configured around the client’s own therapeutic hypothesis, study outlines and representative datasets are available upon request.

Why Choose Alfa Cytology?

Multiple myeloma modeling is really two experiments in one—tumor burden and bone destruction—and a CRO that tracks only the first tells half the story. Alfa Cytology’s integrated imaging, biomarker, and bone-analysis pipeline keeps both halves of the 5TGM1 / 5TGM1-luc dataset complete and aligned.

• Authenticated 5TGM1 / 5TGM1-luc banks with mycoplasma verification and CD138 identity confirmation.
• Syngeneic C57BL/KaLwRij expertise preserving the immune context that drives immunotherapy response.
• Weekly IVIS imaging paired with serum IgG2b ELISA as a dual, corroborating burden readout.
• Endpoint microCT and DEXA bone analysis quantifying the osteolytic lesions that define myeloma bone disease.
• Flexible hosts (C57BL/KaLwRij, NSG, NIH-III nude) matched to each study’s timeline and modality.
• A dedicated project team owning the study from inoculation through the final consolidated data package.

Contact Us

Whether the target is tumor burden, bone protection, or both, Alfa Cytology can configure a 5TGM1 / 5TGM1-luc orthotopic myeloma study that captures the full picture of the disease. Contact us today to discuss study design and imaging cadence, or reach out to our team for a customized quotation.

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