RL95-2 Xenograft Model Service for Endometrial Cancer

The RL95-2 xenograft model offers a robust preclinical platform for evaluating therapeutic efficacy against endometrial carcinoma, leveraging a well-characterized adenosquamous cell line with preserved hormone receptor expression and documented tumorigenic potential in immunodeficient hosts. At Alfa Cytology, we specialize in constructing and validating RL95-2 xenograft models under rigorous quality-controlled conditions, providing pharmaceutical and biotechnology partners with reliable tumor-bearing cohorts, comprehensive endpoint analysis, and tailored study designs to accelerate preclinical development programs in gynecological oncology.

Overview of RL95-2 Xenograft Model for Endometrial Cancer

The RL95-2 cell line was originally established in 1980 from a grade 2 moderately differentiated adenosquamous carcinoma of the endometrium obtained from a 65-year-old patient at the University of Arizona Medical Center. This epithelial cell line retains both cytoplasmic and nuclear estrogen receptor positivity, exhibits a doubling time of approximately 22-34 hours, and demonstrates characteristic glandular dome formation with well-defined junctional complexes and surface microvilli. Genomically, RL95-2 harbors a PTEN frameshift termination mutant (T321fs-ter), an in-frame TP53 deletion (V218del), and PIK3R1/ARID1A mutations, while presenting a microsatellite instability-low (MSI-Low) and copy-number-high profile that aligns most closely with TCGA serous-like cluster characteristics. These molecular and phenotypic attributes render RL95-2 a representative model for investigating hormone-responsive endometrial malignancies and testing targeted agents directed at the PI3K/AKT and p53 pathways.

In vivo, RL95-2 cells form progressively growing tumors in immunocompromised murine hosts, making the xenograft system suitable for pharmacokinetic/pharmacodynamic studies, combination regimen screening, and biomarker discovery. The model supports subcutaneous and orthotopic implantation routes, with subcutaneous engraftment allowing convenient longitudinal tumor volume monitoring and orthotopic implantation recapitulating anatomically relevant tumor-stroma interactions. Tumor latency, growth kinetics, and histopathological features remain consistent across batches when standardized inoculum densities and host strains are employed, ensuring reproducible data generation for multi-arm preclinical trials.

Figure 1. Effect of docosahexaenoic acid (A) or triacsin c (B) on cell viability in RL95-2 endometrial cancer cells. (Chung, Soo-Ho, et al., 2021)

Cell Line Information: RL95-2

RL95-2 is a human endometrial carcinoma cell line with well-documented origin, morphology, and molecular features. The following table summarizes key characteristics relevant to preclinical xenograft application:

Our Services

Alfa Cytology maintains authenticated RL95-2 master and working cell banks under cGMP-aligned cryopreservation protocols, ensuring genetic stability and mycoplasma-free status for every xenograft study. Our experienced team manages the entire model lifecycle---from cell expansion and quality control to inoculation, monitoring, and necropsy---delivering turnkey solutions that meet the stringent reproducibility demands of IND-enabling preclinical programs.

Workflow of RL95-2 Xenograft Model Construction

Construction of the RL95-2 xenograft model follows a standardized, multi-stage workflow designed to ensure batch-to-batch consistency, regulatory compliance, and scientific rigor. Each stage incorporates quality checkpoints to verify cell viability, sterility, and tumorigenic potential before advancing to the next phase.

1. Step 1: Cell Line Resuscitation and Expansion: Authenticated RL95-2 vials are thawed and expanded in complete growth medium under controlled atmospheric conditions (37 degrees C, 5% CO2). Cells are passaged at subconfluence to maintain logarithmic growth and prevent senescence-associated phenotypic drift.
2. Step 2: Pre-Inoculation Quality Control: Expanded cultures undergo comprehensive QC testing, including STR profiling for identity verification, mycoplasma PCR detection, and sterility screening. Only cultures meeting predetermined viability thresholds (>95%) and passing all QC assays proceed to inoculum preparation.
3. Step 3: Inoculum Preparation: Cells are harvested during exponential growth phase using standardized dissociation protocols, washed in serum-free medium, and resuspended at a defined concentration (typically 1x10^7 cells per 100-200 microL) in a supportive matrix such as Matrigel or basement membrane extract to enhance initial engraftment.
4. Step 4: Animal Preparation and Implantation: Immunodeficient recipient mice (commonly NOD scid gamma or athymic nude strains) are acclimatized under pathogen-free conditions. RL95-2 cell suspensions are implanted subcutaneously into the flank or orthotopically into the uterine horn, depending on study objectives, using aseptic surgical technique.
5. Step 5: Post-Implantation Monitoring: Animals are monitored daily for general health, body weight, and palpable tumor formation. Subcutaneous tumors are measured with digital calipers twice weekly; tumor volume is calculated using the modified ellipsoid formula (length x width^2 x 0.5). Orthotopic lesions are monitored by non-invasive imaging where protocol-appropriate.
6. Step 6: Endpoint Analysis and Sample Collection: Upon reaching predetermined endpoint criteria (tumor volume ~1,500-2,000 mm^3 or study duration limit), animals are humanely euthanized. Tumors are excised, weighed, photographed, and processed for histopathology, immunohistochemistry, flow cytometry, or molecular profiling as specified in the study protocol.
7. Step 7: Data Compilation and Reporting: All raw data---including tumor growth curves, body weight trajectories, survival metrics, and histology images---are compiled into a comprehensive study report with statistical analysis, enabling direct integration into regulatory submission packages or peer-reviewed publications.

Figure 2: Schematic workflow illustrating the derivation and construction of the RL95-2 Xenograft Model at Alfa Cytology.

Case Study-RL95-2 Xenograft Model Development

In a representative development project, RL95-2 cells were expanded from authenticated ATCC stocks and implanted subcutaneously into immunodeficient mice to establish a baseline tumor growth profile. The resulting model demonstrated consistent engraftment rates, predictable exponential growth kinetics, and histological preservation of adenosquamous features upon hematoxylin-eosin staining. Pharmacological intervention with a PI3K pathway inhibitor produced dose-dependent tumor growth delay compared with vehicle-treated controls, validating the model's responsiveness to targeted therapy. These preliminary findings support the utility of the RL95-2 xenograft platform for subsequent compound screening, combination regimen evaluation, and biomarker correlation studies under customized protocol designs.

Why Choose Alfa Cytology?

Alfa Cytology combines scientific expertise, operational excellence, and client-centric flexibility to deliver RL95-2 xenograft models that meet the highest preclinical standards. Our integrated service model minimizes handoff delays and ensures seamless progression from study design to final report.

• Authenticated cell lines with verified STR profiles and documented passage history eliminate genetic drift concerns.
• Rigorous mycoplasma and sterility screening at multiple checkpoints safeguards model integrity and data reliability.
• Customizable implantation routes (subcutaneous, orthotopic, or patient-derived xenograft co-implantation) align with diverse study objectives.
• Real-time tumor monitoring and digital data capture enable adaptive decision-making during active studies.
• Comprehensive endpoint analysis spanning histopathology, immunohistochemistry, flow cytometry, and molecular profiling is available in-house.
• Dedicated project scientists provide weekly progress updates and rapid response to protocol amendments.
• Competitive timelines and transparent pricing support efficient budget planning for early-stage and late-stage preclinical programs.

Contact Us

Whether you are initiating a first-in-class endometrial cancer program or advancing a lead compound toward IND submission, our RL95-2 xenograft model service provides the preclinical foundation you need. Reach out to our scientific team today to discuss your specific requirements, receive a customized study proposal, and discover how Alfa Cytology can accelerate your oncology drug development timeline. Please reach out to us today via our inquiry form or email to learn more about our RL95-2 Xenograft Model services.

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