Basic Information
Description
The mouse Ccl2 gene was replaced by human CCL2 coding sequence in B-hCCL2 MC38 cells. Human CCL2 can be secreted by B-hCCL2 MC38 cells.
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Targeting strategy
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Gene targeting strategy for B-hCCL2 MC38 cells. The exogenous promoter and human CCL2 coding sequence was inserted to replace part of murine exon 2~3. The insertion disrupts the endogenous murine Ccl2 gene, resulting in a non-functional transcript.
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Protein Expression Analysis
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CCL2 expression analysis in B-hCCL2 MC38 cells by ELISA. Human CCL2 was detected in the supernatant of B-hCCL2 MC38 cells but not wild-type MC38 cells. Mouse Ccl2 was not detected in the supernatant of wild-type MC38 cells. The 2-D07 clone of B-hCCL2 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-hCCL2 MC38 cells. B-hCCL2 MC38 cells (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into C57BL/6N mice (female, 7-week-old, n=5). 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-hCCL2 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.
