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Focusing on cutting-edge technologies and research advancements,
sharing Aniphe BioLab’s achievements in scientific innovation, Professional insights
and practical experience in the fields of laboratory animals and preclinical research.

Construction of Tumor Animal Models: Colorectal Cancer Edition

Colorectal cancer (CRC) It is the third most common malignancy worldwide, posing a serious threat to human life and health, with the highest incidence observed in the 40–50 age group. Therefore, establishing an appropriate animal model of colorectal cancer is of paramount importance for investigating the mechanisms underlying its initiation and progression, as well as for developing novel anti‑tumor therapeutics.

 
Tumor xenograft model

 

Currently, animal models of colorectal cancer xenografts are classified, based on the origin of the graft, into cell line–derived xenografts (CDX) and patient‑derived xenografts (PDX).

 

CDX is a model established by directly implanting tumor cell lines into immunodeficient mice. This human‑derived xenograft model, generated using tumor cell lines, is easy to perform, cost‑effective, and widely used in cancer research.

 

Aniphe BioLab's  Selected Cell Lines
Human-derived HT-29, HCT-116, SW620
Rodent-derived CT26, MC38
 

Another type of PDX model is established by implanting tumor tissue fragments derived from patients with colorectal cancer into immunodeficient mice. These colorectal cancer PDX models faithfully recapitulate the original tumor’s histopathological features, oncogene expression profiles, and responses to therapeutic agents, thereby demonstrating strong clinical predictive value.

Many patients with colorectal cancer die from liver metastases, as the liver is a common site of metastasis for colon cancer. Based on the site of transplantation, it can also be classified into ectopic transplantation, such as splenic injection (for hepatic metastases).

 

Case Study: CT26 Spleen and Liver Metastasis Model by Aniphe BioLab:

Spleen-to-liver metastasis model Modeling method

| Laboratory animals: 5-week-old female BALB/c

| Experimental Protocol:

① After anesthetizing the mice, a mid‑ventral oblique incision was made, and the spleen was gently exteriorized into the peritoneal cavity using tissue forceps onto a saline‑moistened gauze pad. Cells were then slowly injected into the spleen with a syringe; at the injection site, the capsule became distended and whitened.

② After injection in each group, gently reposition the spleen using tissue forceps, then close the incision with surgical sutures. The entire procedure must strictly adhere to aseptic techniques. Following recovery from anesthesia, continue housing under SPF‑grade conditions.

| Experimental Evaluation System:

Weight curve

Mice were weighed 2–3 times per week after vaccination.

In vivo imaging

In vivo imaging was performed weekly after vaccination.

Photo at the finish line

Photographs of the spleen and liver were taken after the experiment.

 

| Experimental Results:

① Weight curve:

Image source: Aniphe BioLab

 

 

② In vivo imaging:

    According to the analysis of in vivo data, fluorescence signals were clearly observed in both the liver and spleen.

 

Image source: Aniphe BioLab

 

③ Finish-line photo:

Photographs taken at the endpoint also reveal prominent masses in the liver and spleen, indicating successful model establishment.

 

 

Image source: Aniphe BioLab

 
 
 

|Precautions

 

0 1  Approximately 10 days after cell injection, the tumors had already exhibited marked metastasis, and the experimental period was short.
 
0 2  This experiment must be conducted under sterile conditions, with animals housed in an SPF‑grade facility, and supplemented with appropriate antibiotics.
 
03  Inject using an appropriate cell count.
 

Splenocutaneous injection (for hepatic metastasis) predominantly induces liver metastases in colorectal cancer. The resulting model exhibits a phenotype that closely resembles the clinical features of postoperative liver metastases from colorectal cancer, making it widely used for investigating the mechanisms of liver metastasis and for screening drugs that inhibit hepatic metastasis.

 

Chemically induced model

 

Currently, commonly used chemical carcinogens for colon cancer include dimethylhydrazine (DMH), azoxymethane (AOM), and the inflammatory agent dextran sulfate sodium (DSS). These agents are relatively easy to prepare, exhibit good reproducibility, can be rapidly and extensively replicated in a short time, and broadly mimic the process of carcinogenesis, making them widely employed in experimental studies.

 

AOM/DSS–induced colorectal cancer model

 

Genetically engineered mice

 

Genetically engineered animal models of colorectal cancer are established by employing molecular biological techniques to edit genes implicated in the development of colorectal cancer, such as introducing exogenous oncogenes or knocking out endogenous tumor suppressor genes.

 

Hereditary Nonpolyposis Colorectal Cancer (HNPCC): HNPCC is caused by inactivation of the APC gene, leading to loss of heterozygosity, and by mutations in mismatch repair genes (MLH1, MLH2, MSH6, PMS1, and PMS2). Homozygous mice with knockout of either the MLH1 or MLH2 gene develop lymphoid neoplasms and are also predisposed to gastrointestinal tumors, making them an excellent model for studying HNPCC.

 

Familial Adenomatous Polyposis (FAP): Dysregulation of the APC-β-catenin-TCF–driven Wnt signaling pathway is the primary mechanism underlying the development of FAP. A point mutation at codon 850 on one allele of the APC gene gives rise to multiple intestinal adenomas, and mice carrying this mutation are known as Min (multiple intestinal neoplasia) mice; they are considered one of the most ideal experimental models for studying familial adenomatous polyposis (FAP).

 

 

Organoid
 
Organoid models are generated by culturing stem cells or organ‑progenitor cells in vitro using 3D culture techniques, with targeted induction through the addition of specific extracellular matrix mimetics and cytokines, thereby yielding cellular tissues that closely resemble the architecture of their tissue of origin. As a novel research platform, organoids have opened new avenues for cancer research and offer unprecedented opportunities for precision oncology.

 

Mouse organoid models cultured on chips exhibit excellent performance. Confocal images of immunofluorescence staining for mouse small intestinal organoids (left) and colonic organoids (right) on day 7. Image source: [2].

 

Summary

Selecting an appropriate colorectal cancer animal model based on the research objectives is of great significance for studying tumor pathogenesis as well as for clinical diagnosis and treatment. Aniphe BioLab offers a variety of colorectal cancer animal models and provides pharmacological and efficacy evaluation services tailored to related disease models. We welcome all customers to contact us for more information!

 

References:

[1] Cheng Shumin. Construction and Characteristics of Human Primary Colorectal Cancer Xenograft Models in C57BL/6 Mice and NSG Mice [D]. Jining Medical University, 2020. DOI: 10.27856/d.cnki.gjnyx.2020.000020.

[2]Cui Y ,Xiao R ,Zhou Y , et al.Establishment of organoid models based on a nested array chip for fast and reproducible drug testing in colorectal cancer therapy[J].Bio-Design and Manufacturing,2022,5(04):674-686.

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