MSC80 cell line (mouse Schwann cell line) that expresses myelin genes PMP22 and P0 was used in this study55 (link). Cells were grown in a DMEM medium (1X) (Gibco Life Technologies Ref: 11960-044) supplemented with 10% fetal bovine serum, (Gibco Life Technologies Ref: 10500-064), 1%Penicillin/Streptomycin (Gibco Life Technologies Ref: 15070-063), 1%Sodium Pyruvate (Thermo Scientific Ref: SH30239.01), 1% l -glutamine 100X, (Life Technologies Ref: 25030-024) and 2.5 µg/ml of Fungizone Amphotericin B, (Gibco Life Technologies Ref: 15290-026). Cells were incubated at 37 °C in a humidified atmosphere containing 5% CO2.
L glutamine 100x
Manufactured by Thermo Fisher Scientific
Sourced in United States, Germany
L-glutamine (100X) is a cell culture supplement used to support cell growth and proliferation in vitro. It provides a concentrated source of the amino acid L-glutamine, which is essential for various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Sodium pyruvate, by Thermo Fisher Scientific (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Fungizone amphotericin b, by Thermo Fisher Scientific (1 mentions)
Dmem medium 1x, by Thermo Fisher Scientific (1 mentions)
Insulin transferrin selenium, by Merck Group (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Non essential amino acids (neaa), by Merck Group (1 mentions)
Proline, by Thermo Fisher Scientific (1 mentions)
β glycerophosphate, by Merck Group (1 mentions)
L ascorbic acid, by Merck Group (1 mentions)
Seakem le agarose, by Lonza (1 mentions)
B 27 supplement 50x, by Thermo Fisher Scientific (1 mentions)
Mem vitamin solution 100x, by Thermo Fisher Scientific (1 mentions)
Sodium pyruvate 100x, by Thermo Fisher Scientific (1 mentions)
Hmscs, by ScienCell (1 mentions)
Advanced dmem f 12 culture medium, by Thermo Fisher Scientific (1 mentions)
Ibitreat μ slide, by Ibidi (1 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
Facsaria 2, by BD (1 mentions)
5 protocols using l glutamine 100x
1
Culturing Mouse Schwann Cells
2
Embryonic Cartilage Explant Culture
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Pregnant females with embryos at embryonic day 17.5 were euthanized and the cranial base growth plate cartilage, including a small amount of adjacent bone, were rapidly harvested into Hank’s Balanced Salt Solution (HBSS) with magnesium and calcium and without phenol red (Life Technologies/Life Technologies). Cartilage explants were maintained briefly in a passively humidified incubator at 37°C and 8% CO2, then explants were placed on a LabTek 35 mm glass-bottom culture dish and embedded in 1% (wt/vol) SeaKem LE agarose (Lonza) in HBSS. After 5 min of gelation, 3 ml of cartilage culture media (MEM alpha medium without phenol red [Life Technologies] supplemented with 50 mg/ml penicillin−streptomycin [Life Technologies], L-Glutamine 100x [Life Technologies], 10 mM B-glycerophosphate [Sigma-Aldrich], 50 µg/ml L(+) ascorbic acid [Sigma-Aldrich], 1 nM dexamethasone [Sigma-Aldrich], 1 mg/ml proline [Thermo Fisher Scientific], 1% antioxidant [Sigma-Aldrich], 1 mM sodium pyruvate [Life Technologies], 1% nonessential amino acids [Sigma-Aldrich], and 1% Insulin−Transferrin−Selenium [Sigma-Aldrich]) was added into the dish.
3
Co-culture of Glioma Stem Cells and MSCs
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Glioma stem cell line, GSC23 was kindly provided by the MD Anderson Cancer Center, University of Texas35 (link). HMSCs were bought from ScienCell Research Laboratories (Sciencell, Cat. #7500, CA, USA). Similar to previously reported methods36 (link), GSC23 cells were transfected with red fluorescence protein (RFP) gene, and HMSCs were transfected with green fluorescence protein (GFP) gene. In another method, GSC23 cells were also transfected with a lentiviral vector GV348 (sequence element: Ubi-MCS-SV40-puromycin) containing LOXP-STOP-LOXP-RFP gene, and MSCs were transfected with a lentiviral vector GV348 containing a CRE enzyme gene. The RFP, GFP and CRE-LOXP lentiviral vectors were packaged by Shanghai Genechem Co., Ltd (Shanghai, China). MSCs were cultured in Mesenchymal Stem Cell Medium (MSCM, Sciencell, Cat. #7501). GSC23 cells were maintained in Dulbecco’s Modified Eagle Medium/F12 (DMEM/F12), containing 20 ng/ml basic fibroblast growth factor (bFGF), 20 ng/ml epidermal growth factor (EGF), B27 supplement (50X), 2 mmol/l L-glutamine (100X), MEM vitamin solution (100X) and 100 mM sodium pyruvate (100X) (all from Gibco, Carlsbad, CA, USA).
4
Exploring the Independent Life Cycle of C. suis
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To check whether the C. suis life cycle can exclusively occur outside of a host cell, 5 × 103 sporozoites were released into fresh Advanced® DMEM/F-12 culture medium (Gibco) supplemented with 5% fetal calf serum (Gibco) and penicillin/streptomycin plus l -glutamine 100x (Gibco) onto a new uncoated ibidi 8-well ibiTreat®μ-slide (ibidi, Gräfelfing, Germany) and were incubated at 40°C under 5% CO2. The development of parasite stages was monitored daily.
5
Generation of Induced Regulatory T Cells
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Human CD4+CD25loCD45RAhi naive T cells were flow sorted from the PBMCs of healthy donor using BD FACSAria II (BD Bioscience). Cells were differentiated into induced Treg (iTreg) using anti-CD3/CD28 DynaBeads at a 1:4 bead-to-cell ratio in cell culture media [X-VIVO (Cat# 04–418Q, Lonza), supplemented with 10% FBS (Cat# 10100147C, GIBCO), 1% L-Glutamine-100X (Cat# 335050061, GIBCO), 1% MEM Non-Essential Amino Acids-100X (Cat# 11140050, GIBCO), 1 mM sodium pyruvate (Cat# 11360070, GIBCO), 1% Antibiotic-Antimycotic 100x (Cat# 15240112, GIBCO)],
100 U/mL rhIL-2 (Cat# 200–02, Peprotech) and 5 ng/mL hTGF-β1 (Cat# 7754-BH-100/CF, R&D). After 5 days of culture, cells were characterized by intracellular flow cytometry for Foxp3 expression.
100 U/mL rhIL-2 (Cat# 200–02, Peprotech) and 5 ng/mL hTGF-β1 (Cat# 7754-BH-100/CF, R&D). After 5 days of culture, cells were characterized by intracellular flow cytometry for Foxp3 expression.
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