Sy3200 synergy haps cell sorter

Human NK cells were sorted from splenocytes harvested from control or CD7-KO HSC–engrafted NSG-huIL-15 mice based on expression of human CD45 and CD56, using a SY3200 Synergy HAPS cell sorter (Sony Biotechnology). Control and CD7-KO NK cells were evaluated using IncuCyte Live-cell Analysis system. In total, 5000 UM-SCC-9 cells expressing CBR/GFP were incubated for 2 hours in a 96-well plate and imaged prior to the addition of the NK cells at a 5:1 and 10:1 E:T ratio. Real-time images were captured every 3 hours and up to 72 hours, and they were analyzed using the Incucyte software. Data are presented as green object counts (GFP⁺ UM-SCC-9 tumor cells).

Human T were sorted from splenocytes harvested from control or CD7-KO HSC–engrafted NSGS mice based on expression of human CD45 and CD3, using a MoFlo cell sorter (Beckman Coulter) or a SY3200 Synergy HAPS cell sorter (Sony Biotechnology). Cells were cultured in R10 media with the addition of anti-CD3/CD28 beads at a 3:1 bead/cell ratio, and they were counted every 2 days to maintain cell density less than 1 × 10⁶ cells/mL. Flow cytometry was performed 3 days after activation to assess CD7 expression.