Iscove s modified dulbecco s modified eagle s medium

CLL-like cell lines MEC-1 and JVM-3 were purchased from Cobioer Biological Technology Co., Ltd. (Nanjing, China). Short tandem repeat authentication was conducted by Shanghai Biowing Applied Biotechnology Co., Ltd. (Shanghai, China). MEC-1 was established from the peripheral blood of a 61-year-old Caucasian man with chronic B cell leukemia. JVM-3 was constructed from the peripheral blood of a 73-year-old man with B-prolymphocytic leukemia. Neither of these two cell lines harbored NOTCH1 mutation. The culture medium of MEC-1 consisted of 90% Iscove's modified Dulbecco's Modified Eagle's Medium (Gibco, Invitrogen, Grand Island, NY, USA) and 10% fetal bovine serum (Gibco). JVM-3 was cultured in 90% RPMI 1640 culture medium (Gibco) and 10% fetal bovine serum (Gibco). All the cells were incubated in a 5% CO₂ humidified incubator at 37°C. All experiments were conducted with mycoplasma-free cells.

Human chordoma cell lines (U-CH1 and U-CH2) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and incubated in Iscove’s modified Dulbecco’s modified Eagle’s medium (Invitrogen, Carlsbad, CA, USA) and RPMI-1640 medium (Invitrogen) at a ratio of 1:4, supplemented with 10% fetal bovine serum (FBS, Invitrogen) and 1% penicillin/streptomycin (Beyotime, Haimen, Jiangsu, China). Human embryonic kidney 293T (HEK-293T) cells were also obtained from ATCC and cultured in RPMI-1640 medium containing 10% FBS and 1% penicillin/streptomycin. The incubator was maintained at 37°C and 5% CO₂.

Nucleus pulposus cells were isolated from patients with disc herniation as previously reported [26 (link)].
Three chordoma cell lines, including U-CH2, JHC7 and U-CH1 were purchased from BeNa Culture Collection (Beijing, China). All the cell lines were cultured with Iscove’s Modified Dulbecco’s modified Eagle’s medium (Gibco, Grand Island, New York, USA) and Roswell Park Memorial Institute-1640 medium (Gibco) at the ratio of 4:1 plus 10% fetal bovine serum (FBS; Gibco), 1% 100 units/mL penicillin (Gibco) and 1% 100 μg/mL streptomycin (Gibco).