Macs gmp recombinant human interleukin il 7

PBMCS were isolated by Ficoll-Paque (GE Healthcare, cat no. Cytvia 17-1440-03)
from buffy coats obtained from Sanquin Blood Bank). αβT cells were expanded from PBMCs using CD3/CD28 dynabeads (Thermo Fisher scientific, cat no. 40203D) and (1.7 × 10³ IU/ml of MACS GMP Recombinant Human interleukin (IL)-7 (Miltenyi Biotec, cat no. 130-095-361), and 1.5 × 10² IU/ml MACS GMP Recombinant Human IL-15 (Milteny Biotec, cat no. 130-095-762). HL60, RPMI 8226, and SSC9 stably expressing GFP-luciferase was generated by a previously described retroviral transduction protocol (30 (link)). The plasmid containing the GFP and luciferase transgenes was kindly provided by Jeanette Leusen (UMC Utrecht, Utrecht, Netherlands). The following cell lines were obtained from ATCC between 2010 and 2018, HL60 (CCL-240), RPMI 8226 (CCL-155), SCC9 (CRL-1629) and Daudi (CCL-213). Freestyle 293-F cells (R790-07) were obtained from Invitrogen. ML-1, HL60, RPMI 8226 and Daudi were cultured in RPMI (Gibco, cat no. 12017599), 10% FCS (Bodinco), 1% Pen/Strep (Invitrogen, cat no. 11548876). Freestyle 293-F in Freestyle expression medium (Gibco, cat no. 10319322). SCC9 in DMEM (Gibco, cat no. 31966047) 10% FCS, 1% Pen/Strep.

Patient-derived mononuclear cells obtained by leukapheresis were cryopreserved in freezing medium [sodium chloride (NaCl) 0.9% with 10% dimethylsulfoxide and 5% human albumin (HA)]. The material was thawed at 37°C and mixed with five volumes of leukapheresis thaw medium [X-VIVO 15 chemically defined medium without gentamicin and phenol red (Lonza, Breda, The Netherlands), hereafter, called X-VIVO 15, supplemented with 10% HA]. After washing, cells were resuspended in culture medium with cytokines (X-VIVO 15 medium with 5% human serum), 1.7 × 10³ IU/ml of MACS GMP Recombinant Human interleukin (IL)-7 (Miltenyi Biotec, Bergisch Gladbach, Germany), and 1.5 × 10² IU/ml MACS GMP Recombinant Human IL-15 (Miltenyi Biotec).