Basic Information
Description
The mouse Ror1 gene was replaced by human ROR1 coding sequence in B-hROR1 MC38 cells. Human ROR1 is highly expressed on the surface of B-hROR1 MC38 cells.
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Targeting strategy
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The exogenous promoter and human ROR1 coding sequence was inserted to replace part of murine exon 3 and all of exon 4. The insertion disrupts the endogenous murine Ror1 gene, resulting in a non-functional transcript.
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Protein expression analysis
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ROR1 expression analysis in B-hROR1 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hROR1 MC38 cultures were stained with species-specific anti-ROR1 antibody. Human ROR1 was detected on the surface of B-hROR1 MC38 cells but not wild-type MC38 cells. The 1-G06 clone of B-hROR1 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-hROR1 MC38 cells. B-hROR1 MC38 cells (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into C57BL/6N mice (female, 8-week-old, n=8). 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-hROR1 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-hROR1 MC38 cells were subcutaneously transplanted into C57BL/6 mice (n=8). At the end of the experiment, tumor cells were harvested and assessed for human ROR1 expression by flow cytometry. As shown, human ROR1 was highly expressed on the surface of tumor cells. Therefore, B-hROR1 MC38 cells can be used for in vivo efficacy studies of novel ROR1 therapeutics.
