MDCK EGFP cells (65 (link)) and HDF cells (66 (link)) were cultured in Dulbecco’s modified Eagle’s medium (DMEM)–High Glucose medium (Sigma-Aldrich) supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich), penicillin-streptomycin (100 U ml−1; LifeScience), and 2 mM l -glutamine (LifeScience) and were maintained at 37°C and 5% CO2. MDCK FUCCI cells (37 (link)) were cultured in DMEM–High Glucose medium (Sigma-Aldrich) supplemented with 10% FBS (Sigma-Aldrich) and penicillin-streptomycin (100 U ml−1; LifeScience) and were maintained at 37°C and 5% CO2. In the monolayer experiments, a total of 8 × 104 cells/cm2 were seeded on coated substrates made of PDMS (Dow Corning, USA). In the single-cell experiments, a total of 104 cells/cm2 were seeded on the coated substrates.
High glucose medium
Manufactured by Merck Group
Sourced in Germany, United States, Macao
High glucose medium is a type of cell culture medium that contains a high concentration of glucose as the primary energy source for the cells. It is commonly used to support the growth and proliferation of various cell types, particularly those that have high metabolic requirements or are sensitive to glucose levels.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (2 mentions)
Streptomycin, by Merck Group (2 mentions)
Penicillin, by Merck Group (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Hct116wt, by American Type Culture Collection (1 mentions)
Beas 2b, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Hoechst 33258, by Thermo Fisher Scientific (1 mentions)
Mco 19aic, by Panasonic (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Ephrin b2 fc, by Sino Biological (1 mentions)
Nvp bhg712, by Selleck Chemicals (1 mentions)
β glycerophosphate, by Solarbio (1 mentions)
5 protocols using high glucose medium
1
MDCK EGFP and HDF Cell Culture
2
Simulating Microgravity Effects on Epithelial Stem Cells
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EpSCs were purchased from Beijing Bei Na Chuang Lian Biotechnology Research Institute (Cell identification is shown in Supplementary 1). EpSCs was cultured in DMEM (Thermo Fisher Scientific, Waltham, MA) and divided into the simulated microgravity (SMG) group and normal gravity (NG) group (1 g static cultures). Simulated microgravity conditions were simulated using an RCCS (Facer et al.2005 (link)). The cultured solution is composed of 94% high-glucose medium (Sigma, Burlington, MA), 5% fetal bovine serum (Sigma), and 1% penicillin-streptomycin (100 units/ml penicillin and 100 mg/ml streptomycin, Thermo Fisher Scientific, Waltham, MA). The SMG group and NG group were maintained at 37°C in a humidified 5% CO2 incubator. EpSCs were observed every day, and fresh medium was supplemented every 2 to 3 d.
3
Cell Culture Protocols for Cancer Research
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Human cells lines: colon cancer (HCT-116 wt, HT-29), non-small lung carcinoma (A549), and bronchial epithelial cells (BEAS-2B) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA). Isogenic cell line HCT 116 p53−/− (deleted both p53 alleles) were a gift from Dr. Bert Vogelstein. Other cell lines, namely MCF-7, SJSA-1, U2OS, HepG2, and Hep3B were received from the collection stored at Centre of Biotechnology of Silesian University of Technology. All cells were grown in DMEM with a high glucose medium (Sigma-Aldrich, Taufkirchen, Germany) supplemented with 10% (v/v) inactivated fetal bovine serum (FBS) (EURx, Gdańsk, Poland) and 1% antibiotics (10,000 μg/mL of streptomycin and 10,000 units/ml of penicillin) (Sigma-Aldrich, Taufkirchen, Germany) at 37 °C in humidified 5% CO2. The maximum concentration of DMSO in the culture media was 0.4%.
4
Melanoma Cell Line Culture Protocol
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Cell line B16-F10 was purchased from the Rio de Janeiro Cell Bank (BCRJ 0046) (murine melanoma), and Melan-A (murine normal melanocytes) was generously provided by Prof.
Miriam Galvonas Jasiulionis (UNIFESP) in 2015. Consequently, the authors performed no additional authentication. All cell lines were tested to be mycoplasma-free by indirect staining with Hoechst 33258 (Thermo Fisher Scientific, USA) and were used within 3 months of thawing the frozen stock. Cells were grown in DMEM (Dulbecco's Modified Eagle's medium) high glucose medium (Sigma-Aldrich, USA) pH 7.2, supplemented with 10% fetal bovine serum (Gibco, Invitrogen, USA), 100 U/mL penicillin and 100 µg/mL streptomycin, in a 5% CO 2 atmosphere at 37°C (Panasonic MCO-19AIC, Japan). For experiments, cells were detached, centrifuged (160×g for 10 minutes), and suspended in supplemented DMEM medium. B16-F10 cells (5.26×10 4 cells/cm 2 ) were added to microplates for 24 h for cell adhesion. After, S-nitroso-MSA-CS (20 and 40 µg/mL) was added and incubated for additional 24 h.
Miriam Galvonas Jasiulionis (UNIFESP) in 2015. Consequently, the authors performed no additional authentication. All cell lines were tested to be mycoplasma-free by indirect staining with Hoechst 33258 (Thermo Fisher Scientific, USA) and were used within 3 months of thawing the frozen stock. Cells were grown in DMEM (Dulbecco's Modified Eagle's medium) high glucose medium (Sigma-Aldrich, USA) pH 7.2, supplemented with 10% fetal bovine serum (Gibco, Invitrogen, USA), 100 U/mL penicillin and 100 µg/mL streptomycin, in a 5% CO 2 atmosphere at 37°C (Panasonic MCO-19AIC, Japan). For experiments, cells were detached, centrifuged (160×g for 10 minutes), and suspended in supplemented DMEM medium. B16-F10 cells (5.26×10 4 cells/cm 2 ) were added to microplates for 24 h for cell adhesion. After, S-nitroso-MSA-CS (20 and 40 µg/mL) was added and incubated for additional 24 h.
5
Osteogenic Differentiation of MC3T3-E1 Cells
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MC3T3-E1 cells were maintained in high glucose medium (HyClone, Logan, USA) supplemented with 10% (v/v) fetal bovine serum (FBS; BBI, Shanghai, China), 100 U/ml penicillin and 100 µg /ml streptomycin (HyClone, Logan, USA) and cultured with 5% CO 2 at 37°C. For osteogenic differentiation, the cells were cultured in high glucose medium supplemented with 5% FBS, 100 U/ml penicillin, 100 µg /ml streptomycin, 10 -8 mol/L dexamethasone (Sigma, St, Louis, MO) and 10 mmol/L β-glycerophosphate (Solarbio, Beijing, China). The medium was replaced every two days.
Referring to the past researches, EphrinB2-fc (Sino Biological, Beijing, China) was used to stimulate ephrinB2-Ephb4 signaling in the MC3T3-E1 cells at a concentration of 4 µg/ml 36,37 . NVP-BHG712 (Selleck, Shanghai, China) was used to inhibit Ephb4 signaling, and we used it at a concentration of 200 nM 38 .
Referring to the past researches, EphrinB2-fc (Sino Biological, Beijing, China) was used to stimulate ephrinB2-Ephb4 signaling in the MC3T3-E1 cells at a concentration of 4 µg/ml 36,37 . NVP-BHG712 (Selleck, Shanghai, China) was used to inhibit Ephb4 signaling, and we used it at a concentration of 200 nM 38 .
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