In brief, 200 μL Keratinocyte-Serum Free Medium containing the transfected cholesteatoma keratinocytes (1 × 105 cells) was added to the upper chamber of the transwell (Costar, Cambridge, MA). The bottom of the transwell chamber was supplemented with 600 μL Keratinocyte-Serum Free Medium containing FBS (10%, Sigma). The migrated and invaded cells were fixed with paraformaldehyde (4%, Sigma) and stained with crystal violet (0.25%, Solarbio). The upper chamber of the transwell used in the invasion assay was pre-coated with matrigel (Sigma), but the migration assay did not. An inverted microscope (MTX Lab Systems) at 100× magnification was applied to calculate and photograph the migrated and invaded cells.
Keratinocyte serum free medium
Manufactured by Merck Group
Sourced in Canada
Keratinocyte-Serum Free Medium is a specialized cell culture medium designed to support the growth and maintenance of keratinocytes, which are the predominant cell type in the outermost layer of the skin. This medium is formulated to provide the necessary nutrients and growth factors for keratinocytes to thrive in a serum-free environment, making it suitable for various applications in skin research and tissue engineering.
Automatically generated - may contain errors
Lab products found in correlation
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Crystal violet, by Solarbio (1 mentions)
Matrigel, by Merck Group (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
L ascorbic acid 2 phosphate, by Nacalai Tesque (1 mentions)
Fetal bovine serum (fbs), by ICN Biomedicals (1 mentions)
N acetyl l cysteine, by Nacalai Tesque (1 mentions)
Dulbecco s modified eagle s medium f12 medium, by Merck Group (1 mentions)
2 protocols using keratinocyte serum free medium
1
Transwell Assay for Cholesteatoma Keratinocyte Migration and Invasion
2
Cultivation of Human Gastric Cell Lines
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Human gastric adenocarcinoma (AGS) cells were obtained from ATCC (Manassas, VA) and cultured in Dulbecco's modified Eagle's medium F12 medium (Sigma-Aldrich, St. Louis, MO), supplemented with 10% (v/v) fetal bovine serum (ICN Biomedicals, Aurora, OH) and penicillin/streptomycin (Thermo Fisher Scientific, Waltham, MA). Immortalized human gastric (KMU-CS12) cells were obtained from Applied Biological Materials (Richmond, BC, Canada) and cultured in keratinocyte serum free medium (Sigma-Aldrich, St. Louis, MO), supplemented with 2 mmol/L N-acetyl-Lcysteine (Nacalai Tesque, Kyoto, Japan), 0.2 mmol/L L- ascorbic acid 2-phosphate (Nacalai Tesque), and penicillin/ streptomycin (Thermo Fisher Scientific). Cells were kept at 37 C in an incubator under a humidified condition containing 5% carbon dioxide and then passaged and subcultured to 90% confluence with 0.2% trypsin (w/v) every 2 to 3 days.
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