B-hUPAR MC38

Basic Information

Description

The mouse Upar gene was replaced by human UPAR coding sequence in B-hUPAR MC38 cells. Human UPAR is highly expressed on the surface of B-hUPAR MC38 cells.

Targeting strategy

Gene targeting strategy for B-hUPAR MC38 cells. The exogenous promoter and chimeric CDS containing mouse signal peptide and human UPAR coding sequence was inserted to replace part of mouse Upar. The insertion disrupts the endogenous murine Upar gene, resulting in a non-functional transcript.

Protein Expression Analysis

UPAR expression analysis in B-hUPAR MC38 cells by flow cytometry. Single cell suspensions from B-hUPAR MC38 cultures were stained with species-specific anti-UPAR antibody. Human UPAR was detected on the surface of B-hUPAR MC38 cells but not wild-type MC38 cells. The 5-F05 clone of B-hUPAR MC38 cells was used for in vivo experiments.

Tumor growth curve & Body weight changes

Subcutaneous homograft tumor growth of B-hUPAR MC38 cells. B-hUPAR MC38 cells (5×10⁵) and wild-type MC38 cells (5×10⁵) were subcutaneously implanted into B-hUPAR mice (male, 7-9-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-hUPAR MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

Subcutaneous homograft tumor growth of B-hUPAR MC38 cells. B-hUPAR MC38 cells and wild-type MC38 cells were subcutaneously implanted into B-hIL12A/hIL12B/hIL12RB1/hIL12RB2 mice (male, 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-hUPAR MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

Subcutaneous homograft tumor growth of B-hUPAR MC38 cells. B-hUPAR MC38 cells and wild-type MC38 cells were subcutaneously implanted into C57BL/6 mice (male, 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-hUPAR MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

Protein expression analysis of tumor cells

Tumor cells were harvested and assessed for human UPAR expression by flow cytometry. As shown, human UPAR was highly expressed on the surface of tumor cells. Therefore, B-hUPAR MC38 cells can be used for in vivo efficacy studies of novel UPAR therapeutics.

B-hUPAR MC38 cells were subcutaneously transplanted into B-hIL12A/hIL12B/hIL12RB1/hIL12RB2 mice and C57BL/6 (n=6), and on 25 days post inoculation, tumor cells were harvested and assessed for human UPAR expression by flow cytometry. As shown, human UPAR was highly expressed on the surface of tumor cells. Therefore, B-hUPAR MC38 cells can be used for in vivo efficacy studies of novel UPAR therapeutics.