B-hDLL3 MC38|BioMice百奥动物

Common name	B-hDLL3 MC38	Catalog number	311448
Aliases	delta like canonical Notch ligand 3, SCDO1	Disease	Colon carcinoma
Organism	Mouse	Strain	C57BL/6
Tissue types	Colon	Tissue	Colon

模型验证

Description

The mouse Dll3 gene was replaced by human DLL3 coding sequence in B-hDLL3 MC38 cells. Human DLL3 is highly expressed on the surface of B-hDLL3 MC38 cells.

Application

B-hDLL3 MC38 cells have the capability to establish tumors in vivo and can be used for efficacy studies.

Targeting strategy

Gene targeting strategy for B-hDLL3 MC38 cells. The exogenous promoter and human DLL3 coding sequence was inserted to replace part of murine exon 2 and all of exon 5. The insertion disrupts the endogenous murine Dll3 gene, resulting in a non-functional transcript.

Protein expression analysis

DLL3 expression analysis in B-hDLL3 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hDLL3 MC38 cultures were stained with anti-human DLL3 antibody AMG-757 (in house). Human DLL3 was detected on the surface of B-hDLL3 MC38 cells but not wild-type MC38 cells. The 3-G07 clone of B-hDLL3 MC38 cells was used for in vivo experiments.

Tumor growth curve & Body weight changes

Subcutaneous homograft tumor growth of B-hDLL3 MC38 cells. B-hDLL3 MC38 cells (5x105) and wild-type MC38 cells (5x105) were subcutaneously implanted into C57BL/6 mice (female, 7-week-old). 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-hDLL3 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

Tumor growth curve & Body weight changes

Subcutaneous homograft tumor growth of B-hDLL3 MC38 cells. B-hDLL3 MC38 cells (1x10⁶) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into B-hCD3E mice (female, 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-hDLL3 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

Tumor volume and weight measurements

Note：

Due to the presence of immunogenicity, the expression level of hDLL3 in tumor tissue has been slightly reduced. We suggest enrolling 8-10 mice per group, or inoculating the DLL3 cell line in DLL3-related humanized mice, to evaluate tumor efficacy.

Protein expression analysis of tumor cells

B-hDLL3 MC38 cells were subcutaneously transplanted into B-hCD3E mice (n=6), and on 28 days post inoculation, tumor cells were harvested and assessed for human DLL3 expression by flow cytometry using anti-human DLL3 antibody AMG-757 (in house). As shown, human DLL3 was expressed on the surface of tumor cells. Therefore, B-hDLL3 MC38 cells can be used for in vivo efficacy studies of novel DLL3 therapeutics.

In vivo efficacy of CD3 BsAb

Antitumor activity of bispecific antibody (BsAb) X in B-hCD3E mice. B-hDLL3 MC38 cells were subcutaneously implanted into B-hCD3E mice (female, 8 week-old, n=6). Mice were grouped when the tumor size was approximately 100 mm³, at which time they were treated with BsAb X provided by the client with doses and schedules indicated in panel. (A) Tumor volume changes during treatment. (B) Body weight changes during treatment. As shown in panel A, BsAb X was efficacious in controlling tumor growth in B-hCD3E mice, demonstrating that the B-hCD3E mouse model is a powerful tool for in vivo efficacy study of T cell bispecific antibody. Values are expressed as mean ± SEM.