Experimental design of the in vivo orthotopic mouse model. (A) Example of the 5–6 week orthotopic xenograft model. On the initial day, mice were intravesically treated with UMUC3-luc expressing cells (1.0 × 106 cells) following anti-asialo GM (100 μg/body i.p.) antibody treatment. (B) Flowchart of the experimental design. UMUC3-luc cells (1 × 106) were orthotopically transplanted into 5–6-week-old female SCID mice by intravesical administration on day 0. The transplanted tumors were allowed to establish for 7 days. The mice were then randomized and divided into two groups followed by treatment by intravesical injections of 1 × 107 γδT cells along with 5 μM ZOL or PBS once a week for a total of four injections. (C) Flowchart of the experimental design in combination with gemcitabine. Same as in (B), with intraperitoneal gemcitabine pretreatment once a week for a total of four injections (days 6, 13, 20, and 27).