Basic Information
Description
The human BCMA coding sequence was inserted into the exon 1 of mouse Bcma in B-hBCMA CT26.WT cells. Human BCMA is highly expressed on the surface of B-hBCMA CT26.WT cells.
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Targeting strategy
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Gene targeting strategy for B-hBCMA CT26.WT cells. The exogenous promoter and human BCMA coding sequence was inserted into the exon 1 of mouse Bcma B-hBCMA CT26.WT cells. The insertion disrupts the endogenous murine Bcma gene, resulting in a non-functional transcript.
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Protein expression analysis
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BCMA expression analysis in B-hBCMA CT26.WT cells by flow cytometry. Single cell suspensions from wild-type CT26.WT and B-hBCMA CT26.WT cultures were stained with species-specific anti-BCMA antibody. Human BCMA was detected on the surface of B-hBCMA CT26.WT cells but not wild-type CT26.WT cells. The 2-G01 clone of B-hBCMA CT26.WT cells was used for in vivo experiments.
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Tumor growth curve & Body weight changes
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Subcutaneous homograft tumor growth of B-hBCMA CT26.WT cells. B-hBCMA CT26.WT cells (1×105) and wild-type CT26.WT cells (1×105) were subcutaneously implanted into BALB/c mice (female, 7-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-hBCMA CT26.WT cells were able to establish 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-hBCMA CT26.WT cells were subcutaneously transplanted into BALB/c mice (n=6), and on 18 days post inoculation, tumor cells were harvested and assessed for human BCMA expression by flow cytometry. As shown, human BCMA was highly expressed on the surface of tumor cells. Therefore, B-hBCMA CT26.WT cells can be used for in vivo efficacy studies of novel BCMA therapeutics.