H22 / H22-luc Orthotopic Mouse Model Service for Liver Cancer

The H22 / H22-luc orthotopic mouse model for liver cancer plants a chemically induced murine hepatocarcinoma directly into the hepatic parenchyma of immunocompetent mice, where it grows as a primary intrahepatic tumor with a reproducible course toward abdominal invasion and ascites. As a CRO focused strictly on preclinical oncology, Alfa Cytology offers this syngeneic platform as a full service, spanning authenticated H22 / H22-luc cell supply, intrahepatic surgical implantation, scheduled in vivo bioluminescence imaging, and endpoint histopathological assessment.

Overview of H22 / H22-luc Orthotopic Mouse Model for Liver Cancer

H22, also known as Hepatoma-22, is a murine hepatocellular carcinoma line with one of the longest pedigrees in experimental oncology. Registered in Cellosaurus as CVCL_H613, the line arose in situ from the liver of a C3HA mouse treated with ortho-aminoazotoluene (OAAT), a classic hepatocarcinogenic azo dye, and has since been maintained by serial transplantation for generations. The cells grow in vitro in standard serum-supplemented media and engraft aggressively in syngeneic and histocompatible recipients of H-2^d background, including BALB/c and C57BL/6 mice, without any need for immunodeficiency. This syngeneic character preserves the complete tumor-immune dialogue, which is why H22 has become the workhorse model for hepatocellular carcinoma studies in immunocompetent hosts and the default benchmark against which nanoparticle, oncolytic virus, and immune-modulating strategies are evaluated.

Fig 1. Bioluminescence imaging of H22-Luc orthotopic liver tumors demonstrating enhanced suppression of hepatocellular carcinoma following restoration of hepatic IRF8 expression combined with PD-1 blockade. (Wu, Hongxi, et al., 2022)

In orthotopic use, H22 cells are delivered by laparotomy into the left or large hepatic lobe, most commonly at around 10⁶ cells in a small volume of 30% Matrigel; reported tumor formation rates reach 100%, with palpable surface nodules appearing within about a week and progressive disease marked by abdominal organ adhesion, local invasion, and increasing ascites in advanced stages. Luciferase-tagged derivatives (H22-luc) transform this classic model into a quantitative imaging system: after intraperitoneal D-luciferin administration, whole-animal bioluminescence charts intrahepatic tumor expansion on a weekly or semiweekly schedule, ex vivo liver imaging confirms burden at necropsy, and untreated orthotopic cohorts typically reach humane endpoints within roughly four weeks, leaving a defined therapeutic window for intervention studies. The platform has been validated with reference agents such as sorafenib and is widely used to probe the hepatic immune microenvironment. Figure 1 outlines how this model is assembled and monitored.

Cell Line Information: H22 / H22-luc

H22 has supported hepatocarcinoma research for decades and is exceptionally well documented in the literature. The defining characteristics of the parent line and its luciferase-tagged derivative are summarized below.

Our Services

An H22 / H22-luc engagement at Alfa Cytology runs as one continuous pipeline: verified cell banking, laparotomy-based intrahepatic implantation with disciplined postoperative care, imaging visits on a pre-agreed cadence, and a terminal package linking photon-flux curves to H&E- and IHC-verified tumor pathology.

Workflow of H22 / H22-luc Orthotopic Mouse Model Construction

Building an H22 or H22-luc orthotopic liver tumor is a surgical-modeling exercise that rewards precision: the inoculum, the hepatic injection, and the imaging schedule all shape data quality. The workflow below describes how this platform is executed from cell bank to data package.

1. Cell Bank Verification and Expansion: H22 or H22-luc stocks are authenticated and mycoplasma-screened, then expanded in RPMI-1640-based medium and harvested in logarithmic phase for inoculation.
2. Luciferase Calibration (H22-luc Only): In vitro bioluminescent output per cell is verified so that in vivo photon flux can be read as relative tumor burden.
3. Mouse Preparation and Randomization: Immunocompetent recipients, commonly female or male BALB/c or C57BL/6 mice of 4–8 weeks, are acclimatized and randomized by body weight, with anesthesia and perioperative analgesia per approved protocols.
4. Laparotomy and Hepatic Exposure: Under aseptic conditions the abdominal cavity is opened and the left or large hepatic lobe is gently exteriorized for direct cell delivery.
5. Intrahepatic Inoculation: Approximately 1 × 10⁶ cells in 20 µl of 30% Matrigel/PBS are injected slowly into the hepatic parenchyma; the needle is held briefly on withdrawal to limit reflux, and any leakage is absorbed.
6. Closure and Postoperative Care: The abdominal wall and skin are sutured, and animals recover under warming with daily monitoring until fully ambulatory.
7. Longitudinal Bioluminescence Monitoring: At scheduled intervals, mice receive intraperitoneal D-luciferin (150 mg/kg) and are imaged under isoflurane anesthesia 8–15 minutes later, with signal quantified by region-of-interest analysis to track intrahepatic tumor growth and flag abdominal spread.
8. Endpoint Necropsy and Histopathology: At humane endpoint the liver is resected for ex vivo imaging and weighing, ascites volume is recorded, abdominal organs are surveyed for adhesion and metastatic foci, and tissues are processed for H&E and immunohistochemistry (e.g., Ki67, immune markers).

Fig 2. H22 / H22-luc orthotopic mouse model construction workflow.

Case Study-H22 / H22-luc Orthotopic Mouse Model Development

Alfa Cytology has executed preclinical studies on the H22 / H22-luc orthotopic platform for clients evaluating hepatic tumor control, immune-modulating combinations, and delivery technologies, producing longitudinal bioluminescence records, ex vivo liver quantification, ascites measurements, and histopathology-supported endpoint packages. Study designs and representative datasets are shared with interested clients upon request, as each engagement is configured around the client’s own therapeutic concept.

Why Choose Alfa Cytology?

Orthotopic liver modeling is unforgiving: a single imprecise injection or an inconsistent imaging session can blur the difference between treatment groups. Alfa Cytology’s surgical discipline and imaging calibration keep H22 / H22-luc studies crisp, reproducible, and publication-grade.

• Authenticated, mycoplasma-screened H22 / H22-luc cell banks maintained under standardized conditions.
• Laparotomy-based intrahepatic implantation with disciplined leakage control and postoperative care.
• Calibrated in-house IVIS bioluminescence imaging on pre-agreed schedules for quantitative tumor tracking.
• Immunocompetent syngeneic hosts preserving full tumor-immune interactions for immunotherapy studies.
• Endpoint pathology integrating ex vivo imaging, ascites scoring, and H&E/IHC confirmation.
• Responsive project management from feasibility through final data delivery.

Contact Us

Whether the goal is intrahepatic tumor control, peritoneal spread interception, or hepatic immune microenvironment modulation, Alfa Cytology can build the H22 / H22-luc orthotopic study around your candidate approach. Contact us today to discuss your 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.