Basic Information
Description
The mouse Cd151 gene was replaced by human CD151 coding sequence in B-hCD151 MC38 cells. Human CD151 is highly expressed on the surface of B-hCD151 MC38 cells.
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Targeting strategy
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Gene targeting strategy for B-hCD151 MC38 cells. The exogenous promoter and human CD151 coding sequence was inserted to replace part of murine exon 4-5. The insertion disrupts the endogenous murine Cd151 gene, resulting in a non-functional transcript.
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Protein expression analysis
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CD151 expression analysis in B-hCD151 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hCD151 MC38 cultures were stained with species-specific anti-CD151 antibody. Human CD151 was detected on the surface of B-hCD151 MC38 cells but not wild-type MC38 cells. The 2-G11 clone of B-hCD151 MC38 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-hCD151 MC38 cells. B-hCD151 MC38 cells (5×106) and wild-type MC38 cells (5×105) were subcutaneously implanted into C57BL/6 mice (female, 6-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-hCD151 MC38 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-hCD151 MC38 cells were subcutaneously transplanted into C57BL/6 mice (n=6), and on 35 days post inoculation, tumor cells were harvested and assessed for human CD151 expression by flow cytometry. As shown, human CD151 was highly expressed on the surface of tumor cells. Therefore, B-hCD151 MC38 cells can be used for in vivo efficacy studies of novel CD151 therapeutics.
