HCT 116 Metastatic Tumor Model Service for Colon Cancer

The HCT 116 Metastatic Tumor Model Service for Colon Cancer delivers the most widely used human colorectal carcinoma xenograft, distinguished by microsatellite instability, mismatch-repair deficiency, and a wild-type TP53 background. Alfa Cytology supports every facet of this model — authenticated cell banking, immunodeficient host preparation, subcutaneous and orthotopic implantation, splenic and tail-vein metastasis induction, and full histological and molecular endpoint analysis — giving colon cancer programs a versatile in vivo workhorse.

Overview of HCT 116 Metastatic Tumor Model for Colon Cancer

HCT 116 is a human colorectal carcinoma cell line established in the early 1980s by the group of M. G. Brattain from a primary colon adenocarcinoma of a 48-year-old male patient. The line carries one of the cleanest molecular pedigrees in colorectal cancer research: it is microsatellite instability-high (MSI-H) due to inactivation of the MLH1 mismatch-repair protein, harbors oncogenic KRAS (G13D) and PIK3CA (H1047R) mutations together with a heterozygous CTNNB1 alteration, yet retains wild-type TP53, PTEN, and BRAF — a rare genotype combination that has made HCT 116 the standard system for dissecting Wnt/β-catenin signaling, cell-cycle control, and apoptosis in vitro, and for probing chemoresistance to 5-fluorouracil, irinotecan, and oxaliplatin.

Fig 1. Standardized workflow of the HCT116-luc hepatic metastasis model: construction of HCT116-luc cells via lentiviral transduction, subcutaneous transplantation for tumor amplification, orthotopic re-implantation, intraperitoneal D-luciferin administration, metastatic nodule counting, and bioluminescence imaging. (Zhang, Yajuan, et al., 2023)

In immunodeficient hosts, HCT 116 xenografts are remarkably accommodating: as few as 1 × 10⁵ cells implanted subcutaneously produce palpable tumors within 7–10 days, providing rapid, reproducible efficacy readouts that defined early dose–response relationships for standard colorectal chemotherapeutics and now benchmark MEK, PARP, and autophagy-pathway inhibitors. Beyond the flank, the line supports the full spectrum of metastasis-oriented formats — orthotopic microinjection into the cecal wall recapitulates local invasion and dissemination to every clinically relevant organ site, intrasplenic injection yields reliable liver colonization (as few as 1 × 10⁴ cells producing 100% liver metastasis in NOG mice), tail-vein delivery seeds lung lesions trackable by bioluminescence, and intraperitoneal inoculation models carcinomatosis. Engineered resources — MLH1-restored chromosome-transfer derivatives, doxycycline-inducible knockdown clones, and GFP/luciferase reporter lines — further position HCT 116 as the reference platform for MSI-high colon cancer research in vivo.

Cell Line Information: HCT 116

HCT 116 (also written HCT-116 or HCT116) is among the most extensively used colorectal cancer lines worldwide. Its defining characteristics are summarized below.

Our Services

Alfa Cytology maintains authenticated, mycoplasma-free HCT 116 stocks — including MLH1-restored and reporter-tagged derivatives — and operates established nude, SCID, NOD/SCID, and NOG xenograft colonies with surgical capability for cecal, splenic, and tail-vein procedures. Whether the goal is a rapid subcutaneous efficacy screen or a multi-site metastasis program, each study is randomized, quality-controlled, and reported with full statistical interpretation.

Workflow of HCT 116 Metastatic Tumor Model Construction

Building an HCT 116 study pairs the line’s rapid, reliable tumorigenicity with the specific demands of each implantation route. The workflow below describes how the model is constructed and evaluated:

1. Cell Line Qualification: HCT 116 cells are authenticated by STR profiling, confirmed mycoplasma-free, and expanded at controlled passage in McCoy’s 5a medium; engineered derivatives are verified before use.
2. Host Preparation and Grouping: Female or male immunodeficient mice (athymic nude, SCID, NOD/SCID, or NOG, matched to the study design) are acclimatized and randomly allocated to groups.
3. Route-Specific Implantation: High-viability suspensions are delivered as planned: 2–5 × 10⁶ cells subcutaneously for growth studies, microinjected into the cecal wall for orthotopic modeling, injected into the spleen (about 5 × 10⁵ cells) for liver metastasis, or administered via tail vein (about 1 × 10⁶ luciferase-labeled cells) for lung colonization.
4. Model Establishment and Monitoring: Implant sites or surgical recovery are monitored; subcutaneous tumors typically become palpable within 7–10 days, after which caliper measurements, body weight, and clinical status are recorded on schedule.
5. Randomization and Dosing: Animals are re-randomized once tumors reach the target size (e.g., approximately 150–200 mm³), and treatment with vehicle, reference chemotherapeutics (e.g., 5-FU, irinotecan), or test articles begins.
6. In-Life Follow-Up: Tumor growth and tolerability are tracked throughout; luciferase-labeled cohorts undergo serial bioluminescence imaging in vivo to monitor metastatic burden.
7. Terminal Endpoint Analysis: At necropsy, primary tumors are excised and weighed; liver, lungs, peritoneum, and other organs are examined macroscopically and by hematoxylin and eosin histology, with tissues processed for immunohistochemistry or molecular assays as specified.
8. Data Analysis and Reporting: Growth-inhibition metrics, metastatic incidence and burden, and molecular data are statistically analyzed and consolidated into a final study report.

Fig 2. HCT 116 Metastatic Tumor Model construction workflow.

Case Study-HCT 116 Metastatic Tumor Model Development

In a recent preclinical collaboration, Alfa Cytology implemented the HCT 116 metastatic colon cancer model for a sponsor evaluating a candidate therapeutic. The scope included expansion and authentication of cells, establishment of tumors in immunodeficient hosts by the route selected for the program, balanced allocation of vehicle and treatment arms, scheduled monitoring of tumor growth and animal condition, and terminal examination of primary tumors and metastasis-relevant organs with histological confirmation — producing an integrated evidence package that informed the client’s preclinical development decisions. Case formats can be arranged around single-agent efficacy, combination regimens, orthotopic or splenic metastasis quantification, or mechanism-of-action endpoints.

Why Choose Alfa Cytology?

Alfa Cytology brings both the model resources and the technical breadth required to unlock the full utility of this versatile colorectal xenograft. Reasons sponsors select our HCT 116 service include:

• Demonstrated experience with HCT 116 across nude, SCID, NOD/SCID, and NOG host systems.
• Complete route coverage — subcutaneous, orthotopic cecal, intrasplenic, tail-vein, and intraperitoneal formats.
• Access to MLH1-restored isogenic derivatives for mismatch-repair mechanism studies.
• In-house GFP/luciferase reporter lines with imaging-supported longitudinal tracking in vivo.
• Established chemotherapeutic benchmarks (5-FU, irinotecan, oxaliplatin) for assay validation.
• Rigorous randomization, blinded scoring, and clear statistical reporting on agreed timelines.

Contact Us

Ready to put the reference MSI-high colon cancer model to work in your program? The HCT 116 specialists at Alfa Cytology are standing by to design the right study for you. Contact us today for a customized proposal, or reach out to discuss routes, endpoints, and timelines — we look forward to contributing to your research.

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