After 24 h of apigenin treatment, the cells were collected and resuspended at a density of 2 × 103 cells/mL in DMEM/F-12 (Hyclone) supplemented with 20 μg/mL basic fibroblast growth factor (PeproTech, NJ, USA), 20 ng/mL epidermal growth factor (PeproTech), 5 μg/mL insulin (Solarbio), 2% B27 (Gibco), and 10 μg/mL transferrin (Solarbio). Then, 2 mL of cells were seeded into ultralow adherent 6-well dishes (Corning, NY, USA) and cultured at 37°C in 5% CO2 for seven days. Tumorspheres were observed using an inverted fluorescence microscope (Leica).
Ultralow adherent 6 well dishes
Manufactured by Corning
Sourced in United States
Ultralow adherent 6-well dishes are specialized cell culture plates designed to minimize cell attachment. These dishes feature a hydrophilic, non-tissue culture-treated surface that inhibits cell adhesion, allowing for the maintenance of cell suspension cultures and the cultivation of spheroid or organoid models.
Automatically generated - may contain errors
Lab products found in correlation
Transferrin, by Solarbio (1 mentions)
Inverted fluorescence microscope, by Leica (1 mentions)
Basic fibroblast growth factor (bfgf), by Thermo Fisher Scientific (1 mentions)
Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions)
Insulin, by Solarbio (1 mentions)
Stempro accutase, by Thermo Fisher Scientific (1 mentions)
2 protocols using ultralow adherent 6 well dishes
1
Apigenin-Induced Tumorsphere Formation
2
Gastric Cancer Stem Cell Self-Renewal Assay
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The formation of tumorspheres was analyzed to evaluate the self-renewal ability of
gastric CICs when the single cells are suspended in serum-free medium. In short, gastric
cancer cells suspended in stem cell medium were cultivated in ultra-low adherent 6-well
dishes (Corning Incorporated, Corning, NY, USA). The density of the suspended cells was
5,000 cells/well, and the components of stem cell medium included DMEM/F12, B27
(1×), ITS (1×), EGF, and bFGF (both cytokines were of a concentration of
20 ng/mL). After the cells were cultivated in the stem cells medium for 7 days, the
counting of the number of tumorspheres was carried out under a conventional microscope.
For the acquisition of the second passage tumorspheres, the tumorspheres of the first
passage were washed with PBS and then the washed tumor-spheres were dissociated by the
cell dissociation reagent (StemPro® Accutase®;
Thermo Fisher Scientific). Then, the dissociated tumorspheres were then propagated.
gastric CICs when the single cells are suspended in serum-free medium. In short, gastric
cancer cells suspended in stem cell medium were cultivated in ultra-low adherent 6-well
dishes (Corning Incorporated, Corning, NY, USA). The density of the suspended cells was
5,000 cells/well, and the components of stem cell medium included DMEM/F12, B27
(1×), ITS (1×), EGF, and bFGF (both cytokines were of a concentration of
20 ng/mL). After the cells were cultivated in the stem cells medium for 7 days, the
counting of the number of tumorspheres was carried out under a conventional microscope.
For the acquisition of the second passage tumorspheres, the tumorspheres of the first
passage were washed with PBS and then the washed tumor-spheres were dissociated by the
cell dissociation reagent (StemPro® Accutase®;
Thermo Fisher Scientific). Then, the dissociated tumorspheres were then propagated.
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