Glutamine

HCT-116 cells (ATCC–LGC Standards S.r.L., Italy) were cultured in McCoy’s 5A medium (Euroclone, United Kingdom) supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin (10,000 U/mL penicillin and 10 mg/mL streptomycin), and 200 mM L glutamine (all sourced from Euroclone, United Kingdom).
5-Fluorouracil (5-FU)-resistant HCT-116 cells (HCT-116-5-FU-R) were cultured in DMEM (Euroclone, United Kingdom) supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin (10,000 U/mL penicillin and 10 mg/mL streptomycin), and 200 mM L glutamine (all sourced from Euroclone, United Kingdom), and additionally, the culture medium contained 5-fluorouracil (5-FU, cat. n°F6627, Sigma-Aldrich, St. Louis, MO, United States) at concentrations up to 70 µM.
Cells were maintained in a humidified atmosphere containing 5% CO₂ at 37°C and used at early passages for all experiments. The culture medium was changed every 2–3 days, and cells were split at 70%–80% confluence.

MSCs were obtained from the bone marrow of Sprague-Dawley rats (Envigo) by flushing the femur and tibia diaphysis with 2 mL/bone of alpha MEM with 2 mM Glutamine and antibiotics (100 U/mL Penicillin G and 100 µg/mL Streptomycin Sulfate) (Euroclone). MSCs were cultured in a humidified incubator at 37 °C with 5% CO₂ in alpha-MEM medium plus 2 mM Glutamine, antibiotics and 20% fetal bovine serum (Euroclone).

RI-1, OCI-LY8, and SUDHL-8 human DLBCL cells lines with different genetic backgrounds for relevant genes (Table 1) were purchased from the German Collection of Microorganisms and Cell Cultures (DSMZ GmbH; Braunschweig, Germany).
RI-1 and SUDHL-8 cells were cultured in RPMI 1640 medium (cod. R8758; Sigma Aldrich, St. Louis, MO, USA) supplemented with 10% heated-inactivated FBS (cod. ECS0180L; Euroclone, Milan, Italy), 1% glutamine (cod. G7513; Sigma Aldrich, St. Louis, MO, USA), and 1% penicillin/streptomycin (cod. P0781; Sigma Aldrich, St. Louis, MO, USA).
OCI-LY8 cells were cultured in Minimum Essential Medium (MEM) (cod. M2279; Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% heat-inactivated FBS (cod. ECS0180L; Euroclone, Milan, Italy), 1% glutamine (cod. G7513; Sigma-Aldrich, St. Louis, MO, USA), 1% non-essential amino acids (cod. M7145; Sigma-Aldrich, St. Louis, MO, USA), and 1% PES (cod. P0781; Sigma-Aldrich, St. Louis, MO, USA).
All the cell lines were maintained under standard culture conditions (37 °C, 5% CO₂).

MOLM, NB4, and HL60 cell lines were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). MOLM and NB4 cells were grown in RPMI 1640 medium supplemented with 200 nmol/L Glutamine (EuroClone), 10% inactivated fetal bovine serum (FBS) (Sigma-Aldrich, St. Louis, MO, USA), and 0.1% penicillin/streptomycin. HL60 cells were grown in ISCOVE’s medium supplemented with 200 nmol/L Glutamine (EuroClone, Milan, Italy), 10% inactivated FBS, and 0.1% penicillin/streptomycin. All cell lines were maintained at 37 °C with 5% CO₂.

The MYC-N amplified neuroblastoma (NB) cell line HTLA-230 was provided by Dr. E. Bogenmann (Children’s Hospital Los Angeles, CA) (Corrias et al., 1996 (link)) and cultured in RPMI-1640 medium supplemented with 10% heat-inactivated FCS (Biochrom, Berlin, Germany), 50 mg/ml streptomycin, 50 mg/ml penicillin (Sigma-Aldrich), and 2 mm glutamine (Euroclone). The cells were cultured in a humidified environment (95% air/5% CO₂) at 37°C and were used to generate 3D tumor models.
Peripheral blood mononuclear cells (PBMCs) were obtained from blood of volunteer healthy donors by Ficoll-Hypaque gradients (Sigma Aldrich). NK cells were purified by using the NK-cell isolation kit II (Miltenyi Biotec) and were cultured on irradiated PBMCs in RPMI-1640 supplemented with 10% heat-inactivated FCS, 50 mg/ml streptomycin, 50 mg/ml penicillin (Sigma-Aldrich), 2 mm glutamine (Euroclone), 600 IU/ml rhIL-2 (Proleukin; Chiron, Emeryville, CA) and 0.5% v/v phytohemagglutinin (Gibco, Paisley, United Kingdom). After 10 passages, NK cells were checked for purity (>95%) analyzing classical NK cell markers (Castriconi et al., 2007b (link)).

SKOV3 and OVCAR3 human ovarian cancer cells were cultured in RPMI 1640 medium (cod. R8758; Sigma Aldrich, St. Louis, MO, USA) supplemented with 10% heated-inactivated FBS (cod. ECS0180L; Euroclone, Milan, Italy), 1% glutamine (cod. G7513; Sigma Aldrich) and 1% penicillin/streptomycin (cod. P0781; Sigma Aldrich).
OAW42 human ovarian cancer cells were cultured in MEM medium (cod. M2279; Sigma Aldrich) supplemented with 10% heated-inactivated FBS (cod. ECS0180L; Euroclone), 1% glutamine (cod. G7513; Sigma Aldrich), 1% non-essential amino acids (cod. M7145; Sigma Aldrich) and 1% penicillin/streptomycin (cod. P0781; Sigma Aldrich).
SKOV3-GFP-LC3 cells were established in our laboratory and maintained in culture conditions as parental SKOV3 cells. The cells were transfected with the plasmid encoding for GFP-LC3 (as previously described in [24 (link)]) and clones stably expressing the transgenic chimeric DNA were selected upon two weeks culture in medium containing 1 mg/mL G418 disulfate salt solution (cod. G8168; Sigma Aldrich).

The NIH-OVCAR-3 cell line was obtained from ATCC (cod. HTB-161; American Type Culture Collection; Manassas, VA, USA). The OAW42 cell line was obtained from ECACC (cod. 85073102; European Collection of Authenticated Cell Cultures; Porton Down, Salisbury, UK). The KURAMOCHI cell line was obtained from JCRB Cell Bank (cod. JCRB0098; Japanese Collection of Research Bioresources Cell Bank; Japan). The OVCAR3 and KURAMOCHI human ovarian cancer cell lines were cultured in RPMI 1640 medium (cod. R8758; Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS, cod. ECS0180L; Euroclone, Milan, Italy), 1% glutamine (cod. G7513; Sigma-Aldrich), and 1% penicillin/streptomycin (PES, cod. P0781; Sigma-Aldrich).
The OAW42 human ovarian cancer cell line was cultured in Minimum Essential Medium (MEM) (cod. M2279; Sigma-Aldrich) supplemented with 10% heat-inactivated FBS, 1% glutamine, 1% non-essential amino acids (cod. M7145; Sigma-Aldrich), and 1% PES.
The OVCAR3-GFP-LC3 cells were established in our laboratory [15 (link)] and maintained in culture conditions as the parental cell line.
All the cell lines were maintained under standard culture conditions (37 °C, 5% CO₂).