Basic Information
Description
The mouse Efemp1 gene was replaced by human EFEMP1 coding sequence in B-hEFEMP1 MC38 cells. Human EFEMP1 is highly expressed on the supernatant of B-hEFEMP1 MC38 cells. B-hEFEMP1 MC38 cells have the capability to establish tumors in vivo and can be used for efficacy studies.
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Gene targeting strategy
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Gene targeting strategy for B-hEFEMP1 MC38 cells. The exogenous promoter and human EFEMP1 coding sequence was inserted to replace part of murine exon 4. The insertion disrupts the endogenous murine Efemp1 gene, resulting in a non-functional transcript.
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Protein expression analysis
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EFEMP1 expression analysis in B-hEFEMP1 MC38 cells by western blot. Human EFEMP1 was detected in the supernatant of B-hEFEMP1 MC38 cells but not wild-type MC38 cells. The 2-F12 clone of B-hEFEMP1 MC38 cells was used for in vivo experiments.
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Tumor growth curve and body weight changes
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Subcutaneous homograft tumor growth of B-hEFEMP1 MC38 cells. B-hEFEMP1 MC38 cells (5×106) and wild-type MC38 cells (5×105) were subcutaneously implanted into C57BL/6 mice (female, 9-week-old, n=6). 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-hEFEMP1 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.
