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.
-
Application
-
B-hFN14 MC38 cells have the capability to establish tumors in vivo, which can be used for efficacy studies.
-
Targeting Strategy
-
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
-
Pre-inoculation
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 an anti-human 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.
Post-inoculation
B-hFN14 MC38 cells were subcutaneously implanted into C57BL/6 mice (n=5) Tumor cells were harvested on day 28 post inoculation and assessed for FN14 expression by flow cytometry using species-specific antibodies. As shown, human FN14 was highly expressed on the surface of humanized B-hFN14 MC38 tumor cells, indicating B-hFN14 MC38 cells can be used for in vivo efficacy studies of novel FN14 therapeutics.
-
Tumor growth curve & Body weight changes
-
Tumor growth curve & Body weight changes in C57BL/6N Mice
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/6N mice (female, 8-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-hFN14 MC38 cells were able to establish tumors in vivo, which can be used for efficacy studies.
