Basic Information
Description
The mouse Dll3 gene was replaced by the human DLL3 coding sequence in B-hDLL3 MC38 cells. Human DLL3 is highly expressed on the surface of B-hDLL3 MC38 cells.
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Application
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B-hDLL3 MC38 cells have the capability to establish tumors in vivo, which can be used for efficacy studies.
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Targeting Strategy
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The exogenous promoter and human DLL3 coding sequence was inserted to replace part of murine exon 2 and all of exon 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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Pre-inoculation
DLL3 expression analysis in B-hDLL3 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hDLL3 MC38 cultures were stained with species-specific anti-DLL3 antibodies. Human DLL3 was detected on the surface of B-hDLL3 MC38 cells but not wild-type MC38 cells. The 3-G07 clone of B-hDLL3 MC38 cells was used for in vivo experiments.
Post-inoculation
B-hDLL3 MC38 cells were subcutaneously implanted into C57BL/6 mice (n=5). Tumor cells were harvested on day 32 post inoculation and assessed for human DLL3 expression by flow cytometry. As shown, human DLL3 was expressed on the surface of humanized B-hDLL3 MC38 tumor cells, indicating B-hDLL3 MC38 cells can be used for in vivo efficacy studies of novel DLL3 therapeutics.
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Tumor Growth Curve
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Tumor Growth Curve & Body Weight Changes in C57BL/6N Mice
Subcutaneous homograft tumor growth of B-hDLL3 MC38 cells. B-hDLL3 MC38 cells (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into C57BL/6N 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-hDLL3 MC38 cells were able to establish tumors in vivo, which can be used for efficacy studies.
