Basic Information
Description
The human CLDN6 coding sequence was inserted to replace exon 2 of the mouse Cldn6 gene. Human CLDN6 is highly expressed on the surface of B-hCLDN6 MC38 cells.
-
Application
-
B-hCLDN6 MC38 cells have the capability to establish tumors in vivo, which can be used for efficacy studies.
-
Targeting strategy
-
The exogenous promoter and human CLDN6 coding sequence was inserted to replace exon 2 of the mouse Cldn6 gene. This insertion disrupts the endogenous murine Cldn6 gene, resulting in a non-functional transcript.
-
Protein Expression Analysis
-
Pre-inoculation
CLDN6 expression analysis in B-hCLDN6 MC38 cells by flow cytometry. Single cell suspensions from B-hCLDN6 MC38 cultures were stained with an anti-human CLDN6 antibody. Human CLDN6 was detected on the surface of B-hCLDN6 MC38 cells but not wild-type MC38 cells. The 2-B06 clone of B-hCLDN6 MC38 cells was used for in vivo experiments.
Post-inoculation
B-hCLDN6 MC38 cells were subcutaneously implanted into B-hCLDN6 mice (n=5). Tumor cells were harvested on day 32 post inoculation and assessed for human CLDN6 expression by flow cytometry. As shown, human CLDN6 was highly expressed on the surface of humanized B-hCLDN6 MC38 tumor cells, indicating B-hCLDN6 MC38 cells can be used for in vivo efficacy studies of novel CLDN6 therapeutics.
-
Tumor Growth Curve
-
Tumor Growth Curve & Body Weight Changes in Humanized Mice
Subcutaneous homograft tumor growth of B-hCLDN6 MC38 cells. B-hCLDN6 MC38 cells (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into B-hCLDN6 mice (female, 7-week-old, n=5). (A) Average tumor volume ± SEM and (B) body weight (mean ± SEM) were measured twice a week. Volume was expressed in mm3 using the formula: V=0.5 X long diameter X short diameter2. As shown in panel A, B-hCLDN6 MC38 cells were able to establish tumors in vivo, which can be used for efficacy studies.
