DMEM/F-12

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	A modified Boyden chamber (8‐μm pore filter, 24‐well cell clusters) (Chemotaxicell; Kurabo, Osaka, Japan) coated with type I collagen (Nitta Gelatin, Inc., Osaka, Japan) were used for the chemotaxis assay (Suganuma et al. 2012; Aso et al. 2013). Fibroblasts were cultured on polyacrylamide gels or plastic dishes coated with 10 μg/mL type I collagen in DMEM/F‐12 cell culture medium (Thermo Fisher Scientific) containing 10% FBS for 4 days. The cells were brought to a quiescent state overnight by incubation in DMEM/F‐12 containing 0.1% FBS.

Protocol tips

A modified Boyden chamber (8‐μm pore filter, 24‐well cell clusters) (Chemotaxicell; Kurabo, Osaka, Japan) coated with type I collagen (Nitta Gelatin, Inc., Osaka, Japan) were used for the chemotaxis assay (Suganuma et al. 2012; Aso et al. 2013). Fibroblasts were cultured on polyacrylamide gels or plastic dishes coated with 10 μg/mL type I collagen in DMEM/F‐12 cell culture medium (Thermo Fisher Scientific) containing 10% FBS for 4 days. The cells were brought to a quiescent state overnight by incubation in DMEM/F‐12 containing 0.1% FBS.

Publication protocol

A modified Boyden chamber (8‐μm pore filter, 24‐well cell clusters) (Chemotaxicell; Kurabo, Osaka, Japan) coated with type I collagen (Nitta Gelatin, Inc., Osaka, Japan) were used for the chemotaxis assay (Suganuma et al. 2012; Aso et al. 2013). Fibroblasts were cultured on polyacrylamide gels or plastic dishes coated with 10 μg/mL type I collagen in DMEM/F‐12 cell culture medium (Thermo Fisher Scientific) containing 10% FBS for 4 days. The cells were brought to a quiescent state overnight by incubation in DMEM/F‐12 containing 0.1% FBS. The cells were trypsinized, suspended in 400 μL DMEM/F‐12 containing 0.1% FBS (2 × 105 cells/mL), and placed in the upper chamber. PDGF‐BB (P3201; Sigma‐Aldrich) prepared in 0.1% BSA in phosphate‐buffered saline (PBS) dissolved in DMEM/F‐12 containing 0.1% FBS was inserted in the wells of the lower chamber. After incubation with PDGF‐BB (10 ng/ mL) for 6 h at 37°C, the nonmigrated cells on the upper surface of the filter were scraped off with a cotton‐tipped applicator. The migrated cells were fixed and stained with Diff‐Quik (Sysmex, Kobe, Japan) and mounted on glass slides. Cells in five fields per chamber were counted using an inverted microscope (CKX41; Olympus) with a 10× objective. In the 3 section, “n” refers to the number of experiments performed. Each experimental condition was tested in duplicate. Solvents did not affect cell migration at the concentrations used (0.1%/vol.).

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Mammalian cell culture media HLF using DMEM/F-12 from Thermo Fisher Scientific.

Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for DMEM/F-12 below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Mammalian cell culture media HLF using DMEM/F-12 from Thermo Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.