Basic Information
Description
The mouse Fn14 gene was replaced by the human FN14 coding sequence in B-hFN14 MC38 cells. Human FN14 is highly expressed on the surface of B-hFN14 MC38 cells.
-
Targeting Strategy
-
Gene targeting strategy for B-hFN14 MC38 cells. The exogenous promoter and human FN14 coding sequence was inserted to replace part of murine exon 1 and all of exons 2-4. The insertion disrupts the endogenous murine Fn14 gene, resulting in a non-functional transcript.
-
Protein Expression Analysis
-
FN14 expression analysis in B-hFN14 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hFN14 MC38 cultures were stained with species-specific anti-FN14 antibody. Human FN14 was detected on the surface of B-hFN14 MC38 cells but not wild-type MC38 cells. The 3-G11 clone of B-hFN14 MC38 cells was used for in vivo experiments.
-
Tumor growth curve & Body weight changes
-
Subcutaneous homograft tumor growth of B-hFN14 MC38 cells. B-hFN14 MC38 cells (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into C57BL/6 mice (female, 6-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-hFN14 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.
Subcutaneous homograft tumor growth of B-hFN14 MC38 cells. B-hFN14 MC38 cells (5×105, 1×106, 5×106) and wild-type MC38 cells (5×105) were subcutaneously implanted into B-hCD3E mice (female, 7-week-old). 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-hFN14 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.
Subcutaneous homograft tumor growth of B-hFN14 MC38 cells. B-hFN14 MC38 plus cells and wild-type MC38 cells were subcutaneously implanted into B-h4-1BB mice (female, 9-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. As shown in panel A, B-hFN14 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies. Values are expressed as mean ± SEM.
-
Protein expression analysis of tumor cells
-
B-hFN14 MC38 cells were subcutaneously transplanted into C57BL/6 mice (n=5). At the end of the experiment, tumor cells were harvested and assessed for human FN14 expression by flow cytometry. As shown, human FN14 was highly expressed on the surface of tumor cells. Therefore, B-hFN14 MC38 cells can be used for in vivo efficacy studies of novel FN14 therapeutics.