A total of 500 cells transfected with siRNA for 72 h were seeded into low-adhesion 96-well plates in serum-free epithelial basal medium supplemented with B27 (50×) (GIBCO, Life Technologies Corporation, New York, NY, USA), 20 ng/mL EGF (PeproTech Inc., Cranbury, NJ, USA), and 20 ng/mL FGF (PeproTech Inc., Cranbury, NJ, USA) in each well. Fresh microsphere medium was regularly added for supplementary nutrients. Finally, we counted the spheres under a microscope after incubation for 7 days. These experiments were performed in triplicate.
B27 50
Manufactured by Thermo Fisher Scientific
Sourced in United States
B27 (50×) is a cell culture supplement used in the maintenance and differentiation of neural cells. It provides a defined, serum-free, and animal-component-free formulation that supports the growth and viability of a variety of neural cell types. The B27 supplement is typically used at a 1× final concentration in cell culture media.
Automatically generated - may contain errors
Lab products found in correlation
Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions)
Fibroblast growth factor (fgf), by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Welgene (1 mentions)
Epidermal growth factor (egf), by R&D Systems (1 mentions)
Astrocyte medium, by ScienCell (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Antibiotic antimycotic solution, by Thermo Fisher Scientific (1 mentions)
3 protocols using b27 50
1
Spheroid Formation Assay for Cell Viability
2
Culturing Glioblastoma Stem Cells and Normal Astrocytes
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Normal human astrocytes (NHA) were purchased from ScienceCell Research Laboratories (USA) and cultured in Astrocyte Medium (ScienCell Research Laboratories, USA) supplemented with 10% fetal bovine serum (FBS; Gibco, Thermo Fisher Scientific, USA) and 1% penicillin/streptomycin (Welgene, Korea). GBM patient-derived GSC11 and GSC23 were obtained from the University of Texas MD Anderson Cancer Center.22 (link) GSCs were maintained using neurobasal medium (NBE, serum-free Neurobasal media supplemented with basic FGF and EGF). NBE conditions were as follows: Dulbecco’s modified Eagle’s medium (DMEM)/F12 (Welgene) was supplemented with 2% B27 (50×) (Gibco), 1% penicillin/streptomycin (Welgene), epidermal growth factor (EGF; 20 ng/mL; R&D Systems, USA), and basic fibroblast growth factor (bFGF; 20 ng/mL; R&D Systems). Serum-free conditions were as follows: Dulbecco’s modified Eagle’s medium (DMEM)/F12 (Welgene) was supplemented with 5% UltraGROTM–Advanced Cell Culture Supplement (AventaCell) and 1% penicillin/streptomycin (Welgene).
3
3D In Vitro Gastric Cancer Model
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To generate a 3D in vitro model of GC, harvested tissues were washed with PBS and cut into pieces of 0.8–1.2 mm in diameter. Tumors were transferred into 6-well plates (4–6 tissues per well, with 6 wells for each group). Each well was maintained with 1 mL of culture medium. The full medium was supplemented with advanced DMEM/F12 (Gibco,), B-27 (50×, Gibco), and antibiotic-antimycotic solution (Gibco). The tissues were cultured in a humidified incubator at 37 °C with 5% CO2.
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