The mouse Steap1 gene was replaced by human STEAP1 coding sequence in B-hSTEAP1 MC38 cells. Human STEAP1 is highly expressed on the surface of B-hSTEAP1 MC38 cells.

Gene targeting strategy for B-hSTEAP1 MC38 cells. The exogenous promoter and human STEAP1 coding sequence was inserted to replace part of murine exon 3. The insertion disrupts the endogenous murine Steap1 gene, resulting in a non-functional transcript.

STEAP1 expression analysis in B-hSTEAP1 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hSTEAP1 MC38 #2-G02 cultures were stained with species-specific anti-STEAP1 antibody (in house). Human STEAP1 was detected on the surface of B-hSTEAP1 MC38 cells but not wild-type MC38 cells.

Subcutaneous homograft tumor growth of B-hSTEAP1 MC38 cells. B-hSTEAP1 MC38 cells (1x10⁶, 5x10⁶) were subcutaneously implanted into B-hCD3EDG mice (male and female, 7-8-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-hSTEAP1 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

Cell Line Inoculation Precautions:

The cell inoculation amount is between 1E6-5E6.