B-hIL34 MC38

Basic Information

Description

The mouse Il34 gene was replaced by human IL34 coding sequence in B-hIL34 MC38 cells. Human IL34 is highly expressed on B-hIL34 MC38 cells.

Targeting strategy

Gene targeting strategy for B-hIL34 MC38 cells. The exogenous promoter and human IL34 coding sequence was inserted to replace part of murine exon 3 and all of exon 4 and exon 5. The insertion disrupts the endogenous murine Il34 gene, resulting in a non-functional transcript.

 

Protein expression analysis

IL34 expression analysis in B-hIL34 MC38 cells by ELISA. Single cell suspensions from wild-type MC38 and B-hIL34 MC38 cultures were stained with species-specific anti-IL34 antibody. Human IL34 was detectable in the suspensions of B-hIL34 MC38 cells but not in wild-type MC38 cells. The 1-D08 clone of B-hIL34 MC38 cells was used for in vivo experiments.

Tumor growth curve & Body weight changes

Subcutaneous homograft tumor growth of B-hIL34 MC38 cells. B-hIL34 MC38 cells (5×10⁵) and wild-type MC38 cells (5×10⁵) were subcutaneously implanted into C57BL/6N mice (female, 9-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-hIL34 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

Protein expression analysis of tumor cells

B-hIL34 MC38 cells were subcutaneously transplanted into C57BL/6N mice, and on 28 days post inoculation, tumor cells were harvested and assessed for human IL34 and mouse IL34 expression by ELISA. As shown, human IL34 was highly expressed in the suspensions of tumor cells. Therefore, B-hIL34 MC38 cells can be used for in vivo efficacy studies of novel IL34 therapeutics.