Oncolytic virus–loaded neural stem cells (NSCs) inhibit xenograft growth and prolong survival of mice with orthotopic glioblastoma. 2.5 × 105 U87 cells were injected stereotactically into the right hemisphere of the brains of 9- to 12-week-old nu/nu mice followed by intratumoral (IT) injection of CRAd-S-pk7 alone (2.5 × 107 I.U./animal) or loaded into NSCs (5 × 105 NSC/animal loaded with 50 I.U./NSC, total 2.5 × 107 I.U. viral dose) at 5 days after tumor implantation. The median survival was 56 days in group receiving phosphate-buffered saline (PBS) injection compared with 63 days in the group treated with virus alone and 93 days in the group treated with NSCs loaded with oncolytic virus, n = 9, [log-rank, PBS versus CRAd-S-pk7 (2.5 × 107 I.U.), P = 0.0049; PBS versus ReNcell + CRAd-S-pk7 (5 × 106 I.U.), P < 0.0001; CRAd-S-pk7 versus ReNcell + CRAd-S-pk7 (2.5 × 107 I.U.), P = 0.0007].