Basic Information
Description
The exogenous promoter and human CCL1 coding sequence were inserted into the mouse genome randomly. Human CCL1 can be secreted by B-Tg(hCCL1) MC38 cells, and drive more T cells into the tumor microenvironment in C57BL/6 mice bearing MC38 cells.
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Gene Targeting Strategy
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Gene targeting strategy for B-Tg(hCCL1) MC38 cells. The exogenous promoter and human CCL1 coding sequence were inserted into the mouse genome randomly.
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Protein Expression Analysis
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Pre-inoculation
Human CCL1 expression analysis in B-Tg(hCCL1) MC38 cells. Human CCL1 protein was detected in the supernatant of B-Tg(hCCL1) MC38 cells compared to wild-type MC38 cells using a species-specific ELISA kit. The 2-C12 clone of B-Tg(hCCL1) MC38 cells was used for in vivo experiments.
Post-inoculation
Tumor cells were harvested and assessed for mouse and human CCL1 expression. Using species-specific ELISA kit, murine and human CCL1 were highly expressed in the tumor homogenate. Data was shown as Mean ± SEM and analyzed using one-way ANOVA followed Dunnett test compared with G1. -
Tumor Growth Curve
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Subcutaneous homograft tumor growth of B-Tg(hCCL1) MC38 cells. B-Tg(hCCL1) MC38 cells (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into C57BL/6 mice (female, 7-week-old, n=5). (A) Average tumor volume ± SEM, and (B) body weight (Mean± SEM) were measured twice a week. Volume was expressed in mm3 using the formula: V=0.5 × long diameter × short diameter2. As shown in panel A, B-Tg(hCCL1) MC38 cells were able to establish tumors in vivo, which can be used for efficacy studies.
Subcutaneous homograft tumor growth of B-Tg(hCCL1) MC38 cells. B-Tg(hCCL1) MC38 (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into humanized B-hCCR8 mice (female, 8-week-old, n=7). (A) Average tumor volume ± SEM, and (B) body weight (Mean± SEM) were measured twice a week. Volume was expressed in mm3 using the formula: V=0.5 × long diameter × short diameter2. As shown in panel A, B-Tg(hCCL1) MC38 cells were able to establish tumors in vivo, which can be used for efficacy studies.
