Basic Information
Description
The mouse Cxcr2 gene was replaced by human CXCR2 coding sequence in B-hCXCR2 MC38 cells. Human CXCR2 is highly expressed on the surface of B-hCXCR2 MC38 cells.
-
Targeting strategy
-
The exogenous promoter and human CXCR2 coding sequence was inserted to replace part of murine exon 2 ~ 3’UTR. The insertion disrupts the endogenous murine Cxcr2 gene, resulting in a non-functional transcript.
-
Protein expression analysis
-
CXCR2 expression analysis in B-hCXCR2 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hCXCR2 MC38 cultures were stained with species-specific anti-CXCR2 antibody. Human CXCR2 was detected on the surface of B-hCXCR2 MC38 cells but not wild-type MC38 cells. The 6-F02 clone of B-hCXCR2 MC38 cells was used for in vivo experiments.
-
Tumor growth curve & Body weight changes
-
Subcutaneous homograft tumor growth of B-hCXCR2 MC38 cells. B-hCXCR2 MC38 cells (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into C57BL/6N 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-hCXCR2 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.
-
Protein expression analysis of tumor cells
-
B-hCXCR2 MC38 cells were subcutaneously transplanted into C57BL/6N mice (n=5). At the end of the experiment, tumor cells were harvested and assessed for human CXCR2 expression by flow cytometry. As shown, human CXCR2 was highly expressed on the surface of tumor cells. Therefore, B-hCXCR2 MC38 cells can be used for in vivo efficacy studies of novel CXCR2 therapeutics.
