Cryostor cs10 freezing media

Harvesting of engineered UVC was performed using accutase (StemCell Technologies) and cells were counted using a CellDrop cell counter (De-Novix). Cells were irradiated at a total single dose of 10 Gy, before centrifugation at 300 xg for 10 min followed by resuspension in 100 μL of CryoStor-CS10 freezing media (StemCell Technologies). The UVC preparations for use in non-human primate studies were analyzed for endotoxin levels (Wickham Laboratories Ltd) and absence of mycoplasma (Mycoplasma Experience Ltd).

CD14⁺ monocytes were isolated from a 100-ml human Leukopak collected from healthy donors (STEMCELL Technologies, Cambridge, MA, USA). The peripheral blood sample was washed three times in an EasySep cell separation buffer (STEMCELL Technologies) and centrifuged at room temperature. The supernatant was aspirated, and cells were resuspended at 5 × 10⁷ cells/ml in an EasySep buffer. An EasySep CD14⁺ selection kit (STEMCELL Technologies) was used to isolate monocytes. Isolation cocktail mix and magnetic particles from the kit were added to the cells and purified by magnetic separation. A series of incubation and centrifugation steps followed, resulting in an enriched cell suspension of CD14⁺ monocytes. The cells were counted using Vi-CELL XR (Beckman Coulter, Indianapolis, IN, USA), resuspended in CryoStor CS10 freezing media (STEMCELL Technologies), and aliquoted into 2-ml vials, followed by freezing overnight at −80°C and transferring to liquid nitrogen for long-term storage.