Xenoplantation of HS-MM cells was performed as previously described (9 (link),10 (link)). Briefly, SCID-Beige mice were injected with 2.5x107 HS-MM cells into the aponeuroses of the thighs, subsequently 9 mice were randomly divided to three groups, of three mice. A total of 0.3 ml saline, with or without 1x104 murine or 1.5x106 human NK cells was intravenously injected to SCID-Beige mice two weeks following HS-MM xenoplantation. A total of 8 weeks following xenoplantation, the mice were sacrificed, and the tumor size was measured using a caliper and the following formula: Volume=(width2 x length)/2.
NK cells were harvested from splenic cells of BALB/c mice using a mouse NK cell isolation kit (Stemcell Technologies, Inc.), or collected from peripheral blood samples from a healthy volunteer using negative selection and a NK Cell Isolation kit, MACS system (Miltenyi Biotec, Inc.) according to the manufacturer's protocol. Written informed consent was provided from the healthy volunteer prior to the start of the study. The study was approved by the Committee of Gifu Graduate School of Gifu, Japan (approval no. 27-80, 2020-066).
The isolated human NK cells were cultured in T25 flasks (Thermo Fisher Scientific, Inc.) with 10 ml KBM502 medium (Kohjin-bio, Co., Ltd.) at 37˚C in a humidified incubator with 5% CO2 for seven days.
NK cells were harvested from splenic cells of BALB/c mice using a mouse NK cell isolation kit (Stemcell Technologies, Inc.), or collected from peripheral blood samples from a healthy volunteer using negative selection and a NK Cell Isolation kit, MACS system (Miltenyi Biotec, Inc.) according to the manufacturer's protocol. Written informed consent was provided from the healthy volunteer prior to the start of the study. The study was approved by the Committee of Gifu Graduate School of Gifu, Japan (approval no. 27-80, 2020-066).
The isolated human NK cells were cultured in T25 flasks (Thermo Fisher Scientific, Inc.) with 10 ml KBM502 medium (Kohjin-bio, Co., Ltd.) at 37˚C in a humidified incubator with 5% CO2 for seven days.
