22Rv1 / 22Rv1-luc Orthotopic Mouse Model Service for Prostate Cancer

The 22Rv1 / 22Rv1-luc orthotopic mouse model for prostate cancer is a human xenograft platform in which AR-positive, androgen-independent 22Rv1 tumor cells are surgically implanted into the prostate lobes of immunodeficient mice, recreating organ-confined tumor growth, local invasion, and castration-resistant progression in a clinically faithful microenvironment. Alfa Cytology offers this model as a fully managed preclinical solution — from authenticated 22Rv1 / 22Rv1-luc cell expansion and luciferase-stable banking, orthotopic surgical implantation and castration-induced CRPC induction, to scheduled in vivo bioluminescence imaging, metastasis surveillance, and comprehensive histopathology — with every parameter engineered around the sponsor’s compound, regimen, and endpoint strategy.

Overview of 22Rv1 / 22Rv1-luc Orthotopic Mouse Model for Prostate Cancer

22Rv1 is a human prostatic carcinoma epithelial line with a lineage that directly mirrors the clinical course of advanced disease: it was established from the CWR22R xenograft, itself derived from a primary prostate carcinoma that had advanced to osseous metastasis, after the parental androgen-dependent CWR22 tumor regressed under castration and subsequently relapsed in vivo. The resulting line is androgen independent yet remains androgen receptor (AR) responsive — it expresses functional AR carrying the H874Y mutation, secretes prostate-specific antigen (PSA), and is only weakly growth-stimulated by dihydrotestosterone — making it one of the few available lines that authentically represent castration-resistant prostate cancer (CRPC). Critically, 22Rv1 expresses the AR-V7 splice variant, which lacks the ligand-binding domain and stays constitutively active, a molecular signature closely associated with resistance to androgen signaling inhibitors such as enzalutamide.

Fig 1. In vivo bioluminescence imaging of orthotopic 22Rv1-luc CRPC xenografts (DMSO/ENZ/HG122) with H&E, AR and Ki67 staining and quantification. (Cong, Xiaonan, et al., 2021)

In the orthotopic setting, 22Rv1 cells are surgically inoculated into the dorsolateral prostate lobes of immunodeficient mice, where they re-establish tumors within the native organ architecture and give rise to locoregional and lymphatic spread that subcutaneous flank models cannot reproduce. The luciferase-labeled derivative, 22Rv1-luc, is generated by stable lentiviral transduction of the luc2 reporter, allowing tumor take, growth kinetics, and treatment response to be quantified longitudinally by in vivo bioluminescence imaging after luciferin administration. Because the model tolerates post-engraftment castration, a single study can span the androgen-present and castrate phases in the same animals, while intracardiac inoculation of 22Rv1-luc extends the platform to experimental metastasis of the bone, liver, lung, and brain.

Cell Line Information: 22Rv1 / 22Rv1-luc

The table below summarizes the essential characteristics of the 22Rv1 parental line and its luciferase-labeled derivative 22Rv1-luc. Both are human in origin and are engrafted into immunodeficient hosts; no syngeneic counterpart exists for this xenograft pair.

Our Services

From study design to final pathology report, Alfa Cytology operates the 22Rv1 / 22Rv1-luc orthotopic model as a single integrated program: our teams handle cell-line quality assurance, precision urologic surgery, longitudinal bioluminescence scheduling, castration and dosing logistics, and endpoint analytics, so sponsors receive a coherent, audit-ready data package without coordinating multiple vendors.

Workflow of 22Rv1 / 22Rv1-luc Orthotopic Mouse Model Construction

Construction of the orthotopic model follows published xenograft methodology: luciferase-labeled 22Rv1 cells are expanded under authenticated conditions, formulated in a Matrigel-supported suspension, and surgically implanted into the dorsolateral prostate lobes of immunodeficient mice, after which engraftment is verified and tumor burden is tracked serially. The process comprises the following stages.

1. Cell authentication and banking: 22Rv1 or 22Rv1-luc cells are verified by STR profiling, confirmed mycoplasma-free, and expanded into low-passage working banks under standardized culture conditions.
2. Cell harvest and formulation: Cultures are harvested at optimal confluence, counted and viability-checked, then resuspended at 5 × 10⁵ cells per 30 μL in a 1:1 PBS/Matrigel mixture kept on ice until the moment of injection.
3. Anesthesia and surgical exposure: Male immunodeficient mice are anesthetized with isoflurane, a lower abdominal incision opens access to the pelvic organs, and the prostate with its dorsolateral lobes is gently exposed for inoculation.
4. Orthotopic implantation: The 30 μL cell suspension is slowly deposited into the dorsolateral prostate lobe; the needle is retained briefly so the Matrigel partially sets, limiting reflux and standardizing take rates between animals.
5. Closure and perioperative care: The prostate is repositioned, the abdominal wall and skin are sutured in layers, and animals receive analgesia with post-operative monitoring until fully recovered.
6. Engraftment verification and serial imaging: Luciferin is administered intraperitoneally and bioluminescence is acquired on an IVIS-class system to confirm tumor take, then repeated on a weekly cadence to quantify tumor burden and detect early metastatic signal.
7. Intervention and endpoint analysis: Experimental arms — including post-engraftment castration to induce the CRPC state, enzalutamide or candidate-agent dosing, and combination regimens — are executed per study design; endpoints combine caliper and imaging measurements with necropsy, ex vivo organ bioluminescence, and histopathological assessment (H&E plus AR, Ki67, and marker-specific immunohistochemistry).

Fig 2. 22Rv1 / 22Rv1-luc orthotopic mouse model construction workflow.

Case Study-22Rv1 / 22Rv1-luc Orthotopic Mouse Model Development

Alfa Cytology has executed multiple preclinical programs on the 22Rv1 / 22Rv1-luc orthotopic platform, encompassing single-agent and combination testing in the castration-resistant setting, post-ADT progression monitoring, and metastasis-oriented study designs. Across these programs, orthotopically implanted 22Rv1-luc tumors were reliably established, longitudinally quantified by in vivo bioluminescence, and characterized at endpoint by histopathology and immunohistochemical profiling, providing sponsors with decision-supporting evidence packages. Representative results and study-level details are available upon request.

Why Choose Alfa Cytology?

Alfa Cytology unites cell-line expertise, microsurgical capability, and quantitative imaging analytics in one preclinical ecosystem, so each 22Rv1 / 22Rv1-luc orthotopic study is delivered with reproducibility, regulatory-grade documentation, and scientific depth.

• Focused experience with AR-positive CRPC xenografts, including post-castration progression and AR-V7-associated resistance study designs.
• Integrated surgical, imaging, and histopathology teams operating under a unified quality system from cell banking to final report.
• Quantitative in vivo bioluminescence readouts enabling longitudinal tumor tracking and reduced inter-group variability.
• Adaptable study architecture covering androgen-deprivation timing, combination regimens, and customized biomarker endpoint panels.
• Proactive project communication with defined milestones, transparent progress reporting, and fast scientific feedback loops.

Contact Us

Whether your program targets AR-V7-driven resistance, next-generation anti-androgens, or orthotopic efficacy endpoints, Alfa Cytology can shape the 22Rv1 / 22Rv1-luc orthotopic mouse model around your goals. Reach out to us with your study outline to receive a tailored experimental design, timeline, and quotation — or contact us directly and our team will respond promptly.

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