All experiments used E. coli asparaginase. For in vitro asparaginase sensitivity and combination experiments, we used DMEM medium with 862 mg/L of GlutaMAX (Gibco, Life Technologies, Grand Island, NY) and 10% fetal bovine serum. Treatment was with Elspar® asparaginase (Lundbeck Inc., Deerfield, IL). For neurosphere cultures and assays, DMEM/F12 media (with 365 mg/L of L-glutamine and 7.5 mg/L of L-asparagine; Gemini Bio-Products, Sacramento, CA) was used with a one-time addition of L-glutamine. For glutamine rescue (at 1 and 10 mM) and ASNS PCR related experiments, DMEM without L-asparagine or L-glutamine (Cellgro, Manassas, VA) was used, supplemented with 10% fetal bovine serum. ASNase was from Sigma-Aldrich (St. Louis, MO; for animal treatment) or ProSpec (Ness-Ziona, Israel; for ASNS PCR experiments). Temozolomide (CGeneTech, Indianapolis, IN) was dissolved in DMSO. The same amounts of DMSO (1%) were also used for control and ASNase treatment groups (without temozolomide).
L glutamine
Manufactured by Gemini Bio
Sourced in United States
L-glutamine is a non-essential amino acid that plays a crucial role in various metabolic processes in the body. It is an important component in the production of proteins, and it also contributes to the maintenance of the immune system and the health of the gastrointestinal tract.
Automatically generated - may contain errors
Lab products found in correlation
Streptomycin, by Gemini Bio (15 mentions)
Penicillin, by Gemini Bio (14 mentions)
Fetal bovine serum (fbs), by Gemini Bio (13 mentions)
Penicillin streptomycin, by Gemini Bio (11 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (7 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (5 mentions)
Dmem f12, by Thermo Fisher Scientific (5 mentions)
L glutamine, by Thermo Fisher Scientific (5 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (4 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
Sodium pyruvate, by Corning (3 mentions)
Rankl, by Sino Biological (3 mentions)
Mem non essential amino acids solution, by Thermo Fisher Scientific (2 mentions)
Flp in 3t3, by Thermo Fisher Scientific (2 mentions)
Universal mycoplasma detection kit, by American Type Culture Collection (2 mentions)
Trypsin edta, by Gemini Bio (2 mentions)
Advanced rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Hela cells ccl 2, by American Type Culture Collection (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Cholera toxin, by Merck Group (2 mentions)
41 protocols using l glutamine
1
E. coli Asparaginase Sensitivity Assays
2
Culturing NIH/3T3 and HEK293T Cell Lines
3
Quantifying Nucleic Acid Content
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The Quant-iT ™ Picogreen ® assay kit and the Presto Blue kit were from Life Technologies (Carlsbad, US).
Bovine serum, L-glutamine, penicillin and streptomycin were from Gemini Bio-Products (Sera Laboratories International, Bolney, UK). All other reagents were purchased from Sigma-Aldrich (Poole, UK).
Bovine serum, L-glutamine, penicillin and streptomycin were from Gemini Bio-Products (Sera Laboratories International, Bolney, UK). All other reagents were purchased from Sigma-Aldrich (Poole, UK).
4
Cell Seeding for Functional Assays
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Cells were seeded into a 96-well CellCarrier plate (PerkinElmer, cat. no. 6005550) at a density of 5,000–12000 cells/well for cell response assays, and 3000 cell/well to track mitosis, using phenol red free RPMI (Biochrom, cat. no. F1275) media supplemented with charcoal:dextran stripped FBS (Gemini Bio-Products, cat. no. 100-119) and L-Glutamine (Gemini Bio-Products, cat. no. 400-106). All experiments were performed in 100 μl total volume per well.
5
Propagation of HCoV-229E in MRC-5 Cells
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Human coronavirus 229E (HcoV-229E) (ATCC, Cat. No. VR-740) was added at a multiplicity of infection (MOI) of 1.0 to an approximately 85% confluent T75 flask of MRC-5 cells (ATCC, Cat. No. CCL-171), 48 h after plating. The flask was maintained with DMEM (Fisher Scientific, Cat. No. 11–965-118) supplemented with 5% heat inactivated fetal bovine serum (Gemini Bioproducts, Cat. No. 100–500) and 1% L-Glutamine (Gemini Bioproducts, Cat. No. 400–106), incubated at 37 °C with 5% CO2 [15 (link)]. The cell culture supernatant was harvested at 72 h post infection when 80% cytopathic effect (CPE) was observed.
6
NK-92 Cell Culture and Stimulation
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NK-92 cells were cultured in MyeloCult H5100 medium (STEMCELL) containing penicillin–streptomycin (Gemini Bio-Products) and 100 IU/ml of recombinant human IL-2. K562 and Jurkat cells were cultured in RPMI-1640 medium (Gibco) containing 10% fetal bovine serum (FBS) and penicillin–streptomycin. A549 and HeLa cells were cultured in DMEM (Gibco) containing 10% FBS and penicillin–streptomycin. For stimulation, NK-92 cells were washed with phosphate-buffered saline (PBS) and rested overnight in MyeloCult H5100 without IL-2 and then cultured in MyeloCult H5100 with 50 ng/ml of IL-15, IL-21, IL-15 + IL-21, or without any cytokine for 3 days, washed with PBS, and cultured in Advanced RPMI-1640 (Gibco) containing penicillin–streptomycin and 2 mM l -glutamine (Gemini Bio-Products) continuously without cytokine or with 50 ng/ml of IL-15, IL-21, IL-15 + IL-21 for 2 days.
7
Culturing HT-29 Colon Cancer Cells
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HT-29 human colon cancer cells were cultured in monolayers at 37°C in a 5% CO2 incubator in McCoy’s 5A medium containing penicillin (100 units/mL), streptomycin (100 μg/mL), L-glutamine (2 mM), and 10% FBS (Gemini Bio-Products, Calabasas, CA, USA).
8
Culturing Diverse Mammalian Cell Lines
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Murine cancer cell lines, B16 melanoma, MC38 colon cancer, and panc02 pancreatic cancer have been used often in our previous studies. Other mammalian cell lines, HEK293, HeLa, HepG2, MDA-MB-468, and CV-1, were originally obtained from ATCC (Manassas, VA). All mammalian cells were grown in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 100 U/mL penicillin and 100 µg/mL streptomycin, 2 mM L-glutamine, and 10% fetal bovine serum (FBS) (Gemini Bio-Products, West Sacramento, CA) in an incubator at 37 °C with 5% CO2.
9
Culturing Diverse Human Cell Lines
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MCF10A human mammary epithelial cells were a kind gift from Sabrina Spencer and were cultured in growth medium DMEM:F12 (GIBCO, #11320-033) containing 5% horse serum (GIBCO, catalog number 16050114), 20 ng/ml EGF (PreproTech, AF-100-15), 0.5 mg/mL hydrocortisone (Sigma-Aldrich, H0888-1g), 100 ng/ml cholera toxin (Sigma-Aldrich, C8052-2mg) 10 μg/ml insulin (Sigma-Aldrich, I1882-100mg), 1% penicillin, and 1% streptomycin (both from Life Technologies, catalog number 15140-122) as described previously (Debnath et al., 2003 (link); Soule et al., 1990 (link)). HeLa cells (CCL-2) were purchased from the ATCC and cultured in Dulbecco’s modified Eagle medium containing high glucose and pyruvate (Invitrogen, #11995-073) supplemented with 10% fetal bovine serum (Axenia Biologix, #F001), 1% penicillin, 1% streptomycin, and 4 mM L-glutamine (all from Gemini Bio-Products, #400-110). HEK293T cells were obtained from ATCC (CRL-3216) and cultured in the same medium as HeLa. hTERT RPE-1 cells were obtained from ATCC (CRF-4000) and cultured in DMEM:F12 (GIBCO, catalog number 11320-033), 10% fetal bovine serum (Axenia Biologix, #F001), 0.01 mg/ml hygromycin B (Invitrogen, 10687-010), 1% penicillin, and 1% streptomycin. All cells were maintained at 37°C and 5% CO2 and discarded after passage 25.
10
Ciliation Induction in IMCD3 and RPE1 Cells
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The mouse IMCD3 cell lines used in the study were generated from a parental IMCD3-FlpIn cell line (gift from P.K. Jackson, Stanford University, Stanford, CA). IMCD3-FlpIn cells were cultured in DMEM/F12 (11330–057; Gibco) supplemented with 10% FBS (100–106; Gemini Bio-products), 100 U/ml penicillin-streptomycin (400–109; Gemini Bio-products), and 2 mM L-glutamine (400–106; Gemini Bio-products). The RPE1-hTERT cell line (CRL-4000; ATCC) was cultured in DMEM/F12 supplemented with 10% FBS, 100 U/ml penicillin-streptomycin, 2 mM L-glutamine, and 0.26% sodium bicarbonate (25080; Gibco).
Ciliation was induced by serum starvation in media containing 0.2% FBS for 16–24 h.
Ciliation was induced by serum starvation in media containing 0.2% FBS for 16–24 h.
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