The MOLT-4 cell line was cultured in RPMI-1640 medium supplemented with 50 µg/ml streptomycin, 50 IU penicillin, and 20% FBS at 37°C in a humidified atmosphere of 5% CO2. BOSC23 and 293T cell lines were maintained in Dulbecco’s modified Eagle medium with 10% FBS and antibiotics.
T cell differentiation was performed as previously described (Six et al., 2011 (link)). In brief, CD34+ UCB cells were sorted and cultured on confluent OP9-DL1 in home made α-MEM medium (Invitrogen) supplemented with 20% FBS (Hyclone; Thermo Fisher Scientific) and cytokines (5 ng/ml rhFLT3-L, 10 ng/ml rhSCF, and 2 ng/ml rhIL7; Miltenyi Biotec). DNA was extracted at different days of culture. Cord blood samples, harvested with informed consent, were obtained from Saint-Louis Hospital Cord Blood Bank, which is authorized by the French Regulation Agency (reference TCG/12/R/004).
T cell differentiation was performed as previously described (Six et al., 2011 (link)). In brief, CD34+ UCB cells were sorted and cultured on confluent OP9-DL1 in home made α-MEM medium (Invitrogen) supplemented with 20% FBS (Hyclone; Thermo Fisher Scientific) and cytokines (5 ng/ml rhFLT3-L, 10 ng/ml rhSCF, and 2 ng/ml rhIL7; Miltenyi Biotec). DNA was extracted at different days of culture. Cord blood samples, harvested with informed consent, were obtained from Saint-Louis Hospital Cord Blood Bank, which is authorized by the French Regulation Agency (reference TCG/12/R/004).