Basic Information
Description
The mouse Ccr9 gene was replaced by human CCR9 coding sequence in B-hCCR9 MC38 cells. Human CCR9 is highly expressed on the surface of B-hCCR9 MC38 cells.
-
Targeting strategy
-
Gene targeting strategy for B-hCCR9 MC38 cells. The exogenous promoter and human CCR9 coding sequence was inserted to replace part of murine exon 4. The insertion disrupts the endogenous murine Ccr9 gene, resulting in a non-functional transcript.
-
Protein expression analysis
-
CCR9 expression analysis in B-hCCR9 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hCCR9 MC38 cultures were stained with species-specific anti-CCR9 antibody. Human CCR9 was detected on the surface of B-hCCR9 MC38 cells but not wild-type MC38 cells. The 1-G09 clone of B-hCCR9 MC38 cells was used for in vivo experiments.
-
Tumor growth curve & Body weight changes
-
Subcutaneous homograft tumor growth of B-hCCR9 MC38 cells. B-hCCR9 MC38 cells (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into heterozygous B-hCCR9 mice (female, 8-week-old, n=6). 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-hCCR9 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.
-
Protein expression analysis of tumor cells
-
B-hCCR9 MC38 cells were subcutaneously transplanted into heterozygous B-hCCR9 mice (n=6), and on 49 days post inoculation, tumor cells were harvested and assessed for CCR9 expression by flow cytometry. As shown, human CCR9 was highly expressed on the surface of tumor cells. Therefore, B-hCCR9 MC38 cells can be used for in vivo efficacy studies of novel CCR9 therapeutics.
