Description

The mouse Cd155 gene was replaced by human CD155 coding sequence in B-hCD155 MC38 cells. Human CD155 is highly expressed on the surface of B-hCD155 MC38 cells.

Application

Targeting strategy

Gene targeting strategy for B-hCD155 MC38 cells. The exogenous promoter and human CD155 coding sequence was inserted to replace part of murine exon 2-3. The insertion disrupts the endogenous murine Cd155 gene, resulting in a non-functional transcript.

Protein expression analysis

CD155 expression analysis in B-hCD155 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hCD155 MC38 cultures were stained with species-specific anti-CD155 antibody. Human CD155 was detected on the surface of B-hCD155 MC38 cells but not wild-type MC38 cells. The 3-B8 clone of B-hCD155 MC38 cells was used for in vivo experiments.

Tumor growth curve & Body weight changes

Subcutaneous homograft tumor growth of B-hCD155 MC38 cells. B-hCD155 MC38 cells (1x10⁷) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into B-hTIGIT mice (female, 7-week-old, n=5). Tumor volume and body weight were measured twice a week. (A) Average tumor volume ± SEM. (B)  Body weight (Mean± SEM). Volume was expressed in mm³ using the formula: V=0.5 X long diameter X short diameter². As shown in panel A, B-hCD155 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

Protein expression analysis of tumor cells