The mouse Spn gene was replaced by human SPN coding sequence in B-hSPN MC38 cells. Human SPN is highly expressed on the surface of B-hSPN MC38 cells.

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

B-hSPN MC38 cells were subcutaneously transplanted into C57BL/6 mice (n=6). At the end of the experiment, tumor cells were harvested and assessed for human SPN expression by flow cytometry. As shown, human SPN was highly expressed on the surface of tumor cells. Therefore, B-hSPN MC38 cells can be used for in vivo efficacy studies of novel SPN therapeutics.

Subcutaneous homograft tumor growth of B-hSPN MC38 cells. B-hSPN MC38 cells (2x10⁵) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into C57BL/6 mice (female, 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 mm³using the formula: V=0.5 X long diameter X short diameter². As shown in panel A, B-hSPN MC38 cells were able to establish tumorsin vivo and can be used for efficacy studies.

Cell Line Inoculation Precautions: The cell inoculation amount is between 5E5-1E6.

B-hSPN MC38 tumor cells growth of individual mice. B-hSPN MC38 cells (2x10⁵) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into C57BL/6 mice (female, 9-week-old, n=6). As shown in panel, B-hSPN MC38 cells were able to establish tumorsin vivo and can be used for efficacy studies.