Intranasal Delivery of Therapeutic Neural Stem Cells (NSCs) in Murine Model: A Procedure for Tumor-targeted Delivery of Oncolytic Virus Loaded NSCs into Brain Tumor Bearing Mouse Model

To load human neural stem cells with oncolytic virus, infect the stem cells with 50 conditional replicate of adenovirus viral particles per cell according to standard adenovirus transfection protocols for 2 hours at room temperature with periodic tapping. Then, wash the cells three times with fresh medium to remove any unbound viral particles and suspend the cells in sterile saline for intranasal delivery.

When the cells have been modified as experimentally appropriate, place the anesthetized tumor-bearing animals in the supine position on a clean drape over a heat pad and place a padded pillow under the heads to maintain the nostrils at an upright angle. The pillow material should be either alcohol-wipeable plastic or disposable rolled-up paper towels to avoid cross animal contamination.

When the mice are in position, use a micropipette to dispense 2 microliters of the experimental human neural stem cell population of interest into each nostril of the first mouse, visually confirming aspiration of the droplet before applying the next volume of cells. Repeat the delivery to each mouse in turn. Then, wait 5 minutes and apply the next round of cells. When all four rounds of cells have been administered, place the animals in a recovery chamber with monitoring until full recovery.