# Establishment of Coala: a novel 3D and 2D cancer cell line derived from colorectal cancer liver metastasis

## TL;DR

Researchers developed a new 2D and 3D cancer cell line called Coala from colorectal cancer liver metastasis to improve cancer research models.

## Contribution

The novel Coala cell line is derived from liver metastasis and can be cultured in both 2D and 3D formats.

## Key findings

- Coala forms tumors in mice with features similar to the original tumor and expresses CDX2, SATB2, and CK20.
- Whole exome sequencing revealed pathogenic mutations in genes like C8B, PRKCQ, APC, and TP53.
- Coala expresses proteins Snail, Pdx-1, FoxA2, and E-cadherin, verified by antibody array chip analysis.

## Abstract

Metastatic colorectal cancer is one of the most critical causes of cancer-related dead in patients suffering this type of malignancy. As the liver metastases are found in almost 25–50% of all colorectal cancer patients, there is an urgent need for suitable in vitro and in vivo models. To the best of our knowledge, only a limited number of well-described 2D or 3D organoid/spheroid cell lines originated from human liver metastases was reported for this type of malignancy and submitted to ATCC and other repository centers for general use. Here, we present a novel, well-characterized cell line Coala that was derived directly from liver metastasis of patient with colorectal cancer. Coala cancer cell line was derived by 2D and 3D culture technology and can be, therefore, cultured as standard 2D cell line or as 3D spheroid culture. When transplanted into athymic mice, Coala cell line forms fast growing tumors with histological features similar to original tumor and expressed CDX2, SATB2, and CK20. Whole exome sequencing analysis of Coala cell line showed the presence of pathogenic mutations in C8B, PRKCQ, IRGM, FGFR4, POLR1C, APC and TP53 genes. As verified by antibody array chip analysis, Coala cells express Snail, Pdx-1, FoxA2, and E-cadherin. Our cell line represents a new, well characterized in vitro research tool for cancer research and will be available to researchers in the field via biorepository centers.

The online version contains supplementary material available at 10.1007/s13577-025-01256-1.