• Origin: The MC38 cell line is derived from C57BL/6 murine colon adenocarcinoma cells. The cell line is a commonly used murine model for colorectal carcinoma.
• Background Information: CEACAM5 molecule is also called CEA or CD66e, found on epithelial cells. CD66 plays a role in hemophilic and heterophilic adhesion. Human CD66e is heavily glycosylated GPI anchored protein capable of both homophilic and heterophilic adhesion. Disease relevance: The CD66e may play a role in the process of metastasis of cancer cells. CD66e is found in serum and it is clinically used as a tumor marker for early detection of disease due to its expression in adenocarcinomas.
• Gene targeting strategy: The human CEACEAM5 coding sequence was inserted into the ROSA26 locus in B-hCEACAM5 MC38 cells. Human CEACAM5 were highly expressed on the surface of B-hCEACAM5 MC38 cells.
• Tumorigenicity: Confirmed in C57BL/6JNifdc and humanized CEACAM5 mice.
• Application: B-hCEACAM5 MC38 cells have the capability to establish tumors in vivo and can be used for antibody drugs of efficacy studies.
• Notes:

Inoculated cell lines can be suspended with DMEM stock solution.

Before implementing the project, it is recommended to perform tumor growth experiments. The recommended cell inoculation amount is between 1E5-5E5.

In the experiment, it is necessary to ensure that the number of animals inoculated subcutaneously is at least 1.4 times the actual grouping number.

CEACAM5 expression analysis in B-hCEACAM5 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hCEACAM5 MC38 #1-H05 cultures were stained with anti-CEACAM5 antibody (Biolegend, 342308). Human CEACAM5 were detected on the surface of B-hCEACAM5 MC38 cells but not wild-type MC38 cells.

Subcutaneous tumor growth of B-hCEACAM5 MC38 cells. B-hCEACAM5 MC38 (5x10⁵) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into C57BL/6JNifdc (7-week-old, n=6). Tumor volume and body weight were measured twice a week. (A) Average tumor volume. (B) Body weight. Volume was expressed in mm3 using the formula: V=0.5 X long diameter X short diameter². Results indicated that B-hCEACAM5 MC38 cells were able to establish tumors in vivo and could be used for efficacy studies. Values are expressed as mean ± SEM.

CEACAM5 expression evalsuated on B-hCEACAM5 MC38 cells by flow cytometry. B-hCEACAM5 MC38 cells were subcutaneously transplanted into C57BL/6JNifdc (n=6). Upon conclusion of the experiment, tumor cells were harvested and analyzed with anti-human CEACAM5 antibody (Biolegend, 342308) by flow cytometry. Therefore, B-hCEACAM5 MC38 cells can be used for in vivo efficacy studies evalsuating novel CEACAM5 therapeutics.

Note: In humanized CEACAM5 mice can effectively resolve the issue of two cellular subpopulations (FAQ)