HCC95 Xenograft Model Service for NSCLC

The HCC95 xenograft model is a well-established preclinical platform for evaluating therapeutic strategies against lung squamous cell carcinoma (LUSC), a major histological subtype of non-small cell lung cancer (NSCLC) characterized by distinct molecular drivers and aggressive clinical behavior. Alfa Cytology offers a comprehensive HCC95 xenograft model service designed to accelerate your oncology drug discovery pipeline, providing robust in vivo data from tumor establishment through endpoint analysis with rigorous quality control and regulatory-compliant reporting.
Overview of HCC95 Xenograft Model for NSCLC
Non-small cell lung cancer (NSCLC) accounts for approximately 85% of all lung cancer cases, with lung squamous cell carcinoma (LUSC) representing roughly 25–30% of NSCLC diagnoses. LUSC is strongly associated with a history of smoking and typically arises from the central airways and bronchi, exhibiting distinct molecular profiles compared to lung adenocarcinoma. The HCC95 cell line, originally established from a 65-year-old Caucasian male patient, is a well-characterized model of LUSC that faithfully recapitulates the histopathological and molecular features of this disease subtype, including positive expression of squamous differentiation markers cytokeratin 5 (CK5) and p40. Genomically, HCC95 harbors a homozygous frameshift mutation in TP53 (p.Arg335Valfs*10, c.1002delG), a hallmark alteration frequently observed in LUSC, together with copy number alterations in the PI3K pathway. These molecular characteristics make HCC95 an invaluable preclinical tool for studying squamous lung cancer biology and evaluating novel therapeutic interventions.
Fig 1. LPIAT1 knockdown reduces tumorigenesis and extends survival of mice. (Saliakoura, Maria, et al., 2020)
Cell line-derived xenograft (CDX) models utilizing HCC95 cells provide a reproducible and physiologically relevant preclinical system for assessing tumor growth kinetics, drug efficacy, resistance mechanisms, pharmacokinetics, and biomarker responses. When implanted into immunodeficient mice, HCC95 cells reliably form tumors that recapitulate key histopathological features of human LUSC, including epithelial morphology, keratinization, and squamous marker expression. Notably, HCC95 xenografts have been reported to form cystic, fluid-filled tumors in vivo, a phenotype that mimics certain clinical manifestations of NSCLC and offers unique opportunities to investigate tumor microenvironment interactions and angiogenesis. These models serve as a critical bridge between in vitro screening and clinical translation, enabling researchers to generate high-confidence efficacy data for LUSC-directed agents prior to advancing compounds into more complex development stages.
Cell Line Information: HCC95
The HCC95 cell line is a well-characterized human lung squamous cell carcinoma cell line that serves as a robust foundation for preclinical xenograft studies. Below is a comprehensive summary of its biological and culture characteristics:
| Parameter |
Details |
| Cell Line Name |
HCC95 (HCC-95; Hamon Cancer Center 95) |
| ATCC Catalog No. |
CRL-5802 |
| KCLB No. |
70095 |
| RRID |
CVCL_5137 |
| Species |
Homo sapiens (Human) |
| Tissue of Origin |
Lung |
| Histology |
Non-Small Cell Lung Cancer (NSCLC), Squamous Cell Carcinoma (LUSC) |
| Patient Demographics |
65-year-old male, Caucasian |
| Year Established |
1986 |
| Derived From |
Metastatic pleural effusion |
| Key Driver Mutation |
TP53 p.Arg335Valfs*10 (c.1002delG), homozygous |
| PIK3CA Status |
Wild-type (copy number gain) |
| Additional Alterations |
TSC2 mutation; CDKN2A alterations |
| Squamous Markers |
CK5 positive, p40 positive |
| Cell Morphology |
Epithelial-like, adherent growth, monolayer |
| Doubling Time |
~40.6 hours |
| Culture Medium |
RPMI 1640 supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin |
| Culture Conditions |
37°C, 5% CO₂, 95% relative humidity |
| Biosafety Level |
BSL-1 |
| STR Authentication |
Authenticated by short tandem repeat (STR) profiling |
| Tumorigenicity |
High tumorigenic potential in immunodeficient mice |
| Tumor Formation |
Reliable subcutaneous and orthotopic engraftment; forms cystic tumors in vivo |
| Recommended Inoculum |
5 × 10⁶ to 10 × 10⁶ cells per mouse (subcutaneous) |
| Tumor Latency |
Approximately 7–14 days post-inoculation |
| Tumor Growth Pattern |
Aggressive growth with consistent take rates >90% |
| Drug Sensitivity Profile |
Sensitive to paclitaxel; resistant to GSK3 inhibitors (e.g., CHIR99021) |
Our Services
At Alfa Cytology, we leverage our extensive expertise in preclinical oncology model development to deliver validated HCC95 xenograft studies with rapid turnaround times and comprehensive data packages. Our integrated service platform spans from tumor establishment and in-life monitoring through histopathological and molecular endpoint analysis, ensuring that your LUSC-directed compound receives rigorous, publication-quality evaluation in a clinically relevant NSCLC setting.
Workflow of HCC95 Xenograft Model Construction
The construction of a reliable HCC95 xenograft model follows a standardized, quality-controlled workflow designed to ensure reproducible tumor growth, consistent pharmacological responses, and regulatory-compliant data generation. Each study is initiated with thorough cell line authentication and host animal health screening, followed by systematic tumor implantation, monitoring, and endpoint analysis.
- Cell Line Preparation and Quality Control: HCC95 cells are recovered from cryopreserved stocks and expanded under standardized culture conditions (RPMI 1640 + 10% FBS, 37°C, 5% CO₂). Prior to inoculation, cells undergo mycoplasma testing, STR authentication, and viability assessment to confirm identity and ensure optimal engraftment potential.
- Host Mouse Selection and Acclimatization: Immunodeficient mouse strains—most commonly athymic nude (nu/nu), NOD-SCID, or NSG (NOD-scid IL2Rγnull) mice—are selected based on study objectives and immune requirements. Animals are acclimatized for a minimum of 5–7 days under controlled environmental conditions with health monitoring and body weight baseline recording.
- Tumor Cell Inoculation: Log-phase HCC95 cells are harvested, washed, and resuspended in phosphate-buffered saline (PBS) or PBS/Matrigel mixture (typically 1:1 v/v). For subcutaneous models, 5 × 10⁶ to 10 × 10⁶ cells in a volume of 100–200 µL are injected into the right flank. Orthotopic models involve intrathoracic injection to recapitulate the native tumor microenvironment and metastatic behavior.
- Tumor Growth Monitoring and Randomization: Tumor development is monitored by caliper measurement twice weekly, with tumor volume calculated using the modified ellipsoid formula (V = 0.5 × length × width²). Mice are randomized into treatment groups when tumors reach a palpable volume of 100–200 mm³, ensuring balanced baseline tumor sizes across cohorts. Body weight and clinical signs are recorded concurrently.
- Treatment Administration and In-Life Assessment: Test articles are administered according to the predefined dosing regimen (route, frequency, and duration). Tumor volume and body weight are measured at regular intervals throughout the treatment period. Tumor growth inhibition (TGI), tumor growth delay (TGD), and partial or complete response rates are calculated relative to vehicle-treated controls.
- Endpoint Analysis and Data Reporting: At study termination, tumors are excised, weighed, and processed for downstream analyses. Standard endpoints include hematoxylin and eosin (H&E) histopathology, immunohistochemistry (IHC) for proliferation (Ki-67) and apoptosis (cleaved caspase-3) markers, squamous differentiation markers (CK5, p40), pharmacokinetic/pharmacodynamic (PK/PD) assessment, and biomarker profiling. A comprehensive study report with statistical analysis is delivered to the client.
Fig 2. HCC95 xenograft model construction workflow.
Case Study-HCC95 Xenograft Model Development
In a representative preclinical study, HCC95 cells were successfully engrafted into immunodeficient mice to evaluate the efficacy of a novel therapeutic candidate targeting lung squamous cell carcinoma. Following subcutaneous inoculation, tumors established consistently with a take rate exceeding 90%, reaching the target volume range within 10–14 days. Treatment cohorts received the investigational compound via oral gavage or intraperitoneal injection on a defined schedule, while vehicle controls received the formulation buffer. Tumor growth was monitored biweekly via caliper measurement, and body weights were recorded to assess treatment tolerability. At study endpoint, excised tumors were subjected to comprehensive histopathological and molecular characterization, revealing dose-dependent reductions in tumor burden, decreased Ki-67 proliferation indices, and elevated apoptotic markers. Pharmacokinetic sampling confirmed adequate systemic exposure, and the overall data package supported the compound's advancement into subsequent preclinical development stages. These findings demonstrate the utility of the HCC95 xenograft model as a robust platform for generating translational efficacy data in LUSC.

Why Choose Alfa Cytology?
Alfa Cytology combines scientific rigor, operational efficiency, and client-centric flexibility to deliver preclinical xenograft studies that meet the highest standards of the pharmaceutical and biotechnology industries. Our HCC95 NSCLC xenograft service is designed to accelerate your drug discovery timeline while ensuring data integrity and regulatory readiness.
- Expertise in LUSC models: Our scientific team has deep experience with squamous cell carcinoma biology and understands the nuances of modeling this histologically distinct NSCLC subtype, including TP53-driven tumor progression and PI3K pathway alterations.
- Validated, authenticated cell lines: All HCC95 stocks are STR-authenticated, mycoplasma-negative, and maintained under rigorous quality control to ensure batch-to-batch consistency and reliable tumor engraftment.
- Flexible study design: We accommodate diverse dosing regimens, combination therapy protocols, biomarker-driven endpoints, and custom analytical requirements tailored to your compound's mechanism of action and development stage.
- Comprehensive endpoint portfolio: From standard tumor growth inhibition and histopathology to advanced molecular profiling, PK/PD integration, and biomarker validation, we provide a full spectrum of analytical capabilities.
- Regulatory-compliant operations: Our vivarium and laboratories operate under IACUC-approved protocols with adherence to GLP-like standards, ensuring data packages suitable for IND-enabling and regulatory submissions.
- Rapid study initiation and reporting: Typical HCC95 xenograft studies can be initiated within 2–4 weeks of contract execution, with comprehensive draft reports delivered promptly after study completion.
Contact Us
Ready to advance your lung squamous cell carcinoma therapeutic program with a validated HCC95 xenograft model? Contact us today to discuss your study requirements, receive a customized project proposal, and learn how Alfa Cytology can accelerate your preclinical oncology research. Our team of experienced scientists is standing by to design a study protocol that aligns with your discovery objectives and delivers actionable, high-quality data.
Reference
- Saliakoura, Maria, et al. "The ACSL3-LPIAT1 signaling drives prostaglandin synthesis in non-small cell lung cancer." Oncogene 39.14 (2020): 2948-2960.
For research use only. Not intended for any clinical use.