Dmem hg

TMSCs were isolated and expanded as previously described [9 (link)]. Briefly, we obtained tonsil tissues from patients (<10 years old) undergoing tonsillectomy with informed written consent. This protocol was approved by the Institutional Review Board (ECT-11-53-02) of Ewha Womans University Mokdong Hospital, Korea. Isolated tonsil tissues were mechanically minced with scissors and then enzymatically digested in Dulbecco’s Modified Eagle’s Medium containing high glucose (4500 mg/L) (DMEM-HG; Welgene Inc., Gyeongsan, Korea) supplemented with 10% fetal bovine serum (FBS; Corning, Corning, NY, USA), 100 U/mL penicillin, 100 U/mL streptomycin, 0.25 μg/mL amphotericin B, 210 U/mL collagenase type I (Thermo Fisher Scientific, Waltham, MA, USA), and 10 mg/mL DNase (Sigma–Aldrich, St. Louis, MO, USA) at 37 °C for 30 min. After filtering the digested tissues, mononuclear cells were obtained by density gradient centrifugation using Ficoll-Paque (GE Healthcare, Piscataway, NJ, USA) and were then plated in a T-150 culture flask for 48 h.
Nonadherent cells were discarded, and adherent cells were further incubated in the culture medium (DMEM-HG containing 10% FBS, 100 U/mL penicillin, 100 U/mL streptomycin, and 0.25 μg/mL amphotericin B). TMSCs were used between passages 4–7 for all experiments.