Penicillin streptomycin solution

Human LF cells were isolated from LF tissues as described previously^{13 (link)}. After the LF tissues were obtained from patients, the samples were washed three times with phosphate-buffered saline (PBS, Boster, Wuhan, China). Subsequently, the LF tissues were cut into approximately 0.5 mm³ granules and digested with 0.2% type I collagenase (Sigma, USA) for 1 h at 37 °C in a cell incubator. Then, 0.2% type I collagenase was removed, and the samples were washed with Dulbecco’s modified Eagle’s medium (DMEM, Gibco, USA). Finally, the specimens were incubated with DMEM containing 15% fetal bovine serum (Gibco, USA) and 1% penicillin‒streptomycin solution (Biosharp, Guangzhou, China) in a cell incubator with a 5% concentration of CO₂ at 37 °C. LF cells within four generations were used for subsequent experiments.

Two osteosarcoma cell lines (U2OS and MNNG/HOS) were obtained from the Procell Life Science & Technology Co., Ltd. U2OS and MNNG/HOS were correspondingly cultured in McCoy’s 5A (Procell, China) and MEM (Procell, China) supplemented with 10% fetal bovine serum (FBS, Gibco, USA) and 1% penicillin-streptomycin solution (Biosharp, China) at 37°C with saturated humidity and 5% CO2. The average time of culture medium exchange was 24-48h. The cells were digested with trypsin-EDTA (Gibco, USA) and passaged when cell adhesion exceeded 80% confluency.

Jerusalem artichoke was collected from Jingbian County, Shaanxi Province (China). Fructose standard was purchased from Shanghai Yuanye Biotechnology Co., Ltd. Other chemical regents are analytical grade and purchased from Sinopharm (Shanghai, China). RPMI 1640 (Biological Industries, Kibbutz Beit Haemek, Israel) containing 10% fetal bovine serum (Biological Industries, Kibbutz Beit Haemek, Israel) and 1% Penicillin-Streptomycin solution (Biosharp®, Anhui, China) was utilized as culture solution for Vero cells. Vero cells were cultured in CO₂ incubator (HF90, Lishen Scientific Instrument Co., Ltd, Shanghai, China) at 37°C with a humidified atmosphere containing 5% CO₂. Respiratory syncytial virus (RSV) and Vero cells were provided by collaborative innovation center of antiviral traditional Chinese medicine, Shandong University of traditional Chinese medicine. RSV was stored in medical cryogenic freezer (Haier, Shandong, China) at -80°C.

All GBM cells in this article were obtained from the Tumor Cell Repository of the National Cancer Institute (Frederick, USA). These cells were grown in the optimal conditions of 37 °C, and 5% CO2 in high-Glucose media supplemented with 15% FBS (Hyclone, Australia) and 1% penicillin–streptomycin solution (Biosharp, China). According to the manufacturer's instructions, the Lipofectamine 3000 reagent (ThermoFisher, USA) was employed to transfect 2 ug of the NUDT1 plasmid (GenePharma, China) into cells when the cell density reached roughly 85%. The plasmid sequence of NUDT1 included the forward sequence (GGUUCCAGCUGGAUCAGAUTT) and the reverse sequence (AUCUGAUCCAGCUGGAACCTT).

Human umbilical vein endothelial cells (HUVECs) were purchased from Cell Bank of Chinese Academy of Sciences (Shanghai, China). HUVECs were grown in RPMI 1640 medium (HyClone, Logan, UT, USA) supplemented with 10% fetal bovine serum (Evergreen, Hangzhou, Zhejiang, China) and 1% penicillin–streptomycin solution (Biosharp, Hefei, Anhui, China) and maintained at 37 °C in a 5% CO₂ humidified atmosphere.
Umbilical cord blood was collected from donors with informed consent, which was approved by the Ethics Committee of Huazhong University of Science and Technology. The mononuclear cells were fractionated from the umbilical cord blood by gradient centrifugation with human peripheral blood lymphocyte separation solution (TBD, Tianjin, China) according to the manufacturer’s instructions. The isolated mononuclear cells were seeded onto cell culture dishes coated with 0.01% human fibronectin (Solarbio, Beijing, China) at a density of 5 × 10⁶ cells/cm² in EGM-2 BulletKit (Lonza, Basel, Switzerland), and maintained at 37 °C in a 5% CO₂ humidified atmosphere. EPCs from passages 1 to 3 were used in the subsequent studies.