Basic Information
Description
The mouse Dll3 gene was replaced by chimeric human DLL3 coding sequence in B-hDLL3 B16-F10 cells. Human DLL3 is highly expressed on the surface of B-hDLL3 B16-F10 cells.
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Targeted allele
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Gene targeting strategy for B-hDLL3 B16-F10 cells. The exogenous promoter and chimeric human DLL3 coding sequence was inserted to replace part of murine exon 2 and all of exons 3-5. The insertion disrupts the endogenous murine Dll3 gene, resulting in a non-functional transcript.
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Protein expression analysis
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DLL3 expression analysis in B-hDLL3 B16-F10 cells by flow cytometry. Single cell suspensions from wild-type B16-F10 and B-hDLL3 B16-F10 cultures were stained with species-specific anti-DLL3 antibody. Human DLL3 was detected on the surface of B-hDLL3 B16-F10 cells but not wild-type B16-F10 cells. The 1-E07 clone of B-hDLL3 B16-F10 cells was used for in vivo tumor growth assays. -
Tumor growth curve & Body weight changes
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Subcutaneous homograft tumor growth of B-hDLL3 B16-F10 cells. B-hDLL3 B16-F10 cells (2×105) and wild-type B16-F10 cells (2×105) were subcutaneously implanted into C57BL/6 mice (female, 7-week-old, n=8). Tumor volume and body weight were measured three times 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-hDLL3 B16-F10 cells were able to form tumors in vivo and can be used for efficacy studies.
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Protein expression analysis of tumor cells
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B-hDLL3 B16-F10 cells were subcutaneously transplanted into C57BL/6 mice (n=8), and on 18 days post inoculation, tumor cells were harvested and assessed for human DLL3 expression by flow cytometry. As shown, human DLL3 was highly expressed on the surface of tumor cells. Therefore, B-hDLL3 B16-F10 cells can be used for in vivo efficacy studies of DLL3 therapeutics.
