Mammalian cell culture media HLF

Cells are sourced from various tissues to grow them in in-vitro conditions. Therefore, cell specific nutrients are important for their survival, maintenance and growth. Determining the appropriate cell culture media is a challenge if you are growing a cell line or a microorganism for the first time. Established cell lines, primary cells, stem cells, bacteria and Yeast all require varied nutrients from basic to complex. Based on the cell type, one can easy find what media and nutrients your peers have used before you try to reinvent the wheel.

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FBMTM Fibroblast Growth Basal Medium

Lonza

Upstream tips	Protocol tips	Downstream tips
	NPM was sterilized by ultraviolet light to kill any possible pathogens in the air sample. NPM-protein corona (NPC) samples were prepared as described above and dissolved in the medium by sonication. HLFs (ATCC® PCS-201-013™) were routinely grown using FGM™ fibroblast growth media (Lonza, USA) in a humidified atmosphere of 95% air and 5% CO2 at 37 °C. Cells were suspended at 2 × 106 cells/ml and mixed with the NPC solutions. NPM concentrations of NPC-HLFs were fixed at 25, 50 and 100 μg/ml, respectively. To demonstrate the bioactivity of the protein corona, transforming growth factor beta 1 (Sigma, USA), NPC-vitamin C, pure serum and pure nanoscale PM2.5 (NPM) were also mixed with HLFs following the same procedure described above. The final exposure concentrations were 10 ng/ml (TGF-β1), 0.5 mg/ml (vitamin C) and 100 μg/ml (NPM).

Protocol tips
NPM was sterilized by ultraviolet light to kill any possible pathogens in the air sample. NPM-protein corona (NPC) samples were prepared as described above and dissolved in the medium by sonication. HLFs (ATCC® PCS-201-013™) were routinely grown using FGM™ fibroblast growth media (Lonza, USA) in a humidified atmosphere of 95% air and 5% CO2 at 37 °C. Cells were suspended at 2 × 106 cells/ml and mixed with the NPC solutions. NPM concentrations of NPC-HLFs were fixed at 25, 50 and 100 μg/ml, respectively. To demonstrate the bioactivity of the protein corona, transforming growth factor beta 1 (Sigma, USA), NPC-vitamin C, pure serum and pure nanoscale PM2.5 (NPM) were also mixed with HLFs following the same procedure described above. The final exposure concentrations were 10 ng/ml (TGF-β1), 0.5 mg/ml (vitamin C) and 100 μg/ml (NPM).

Protocol tips

NPM was sterilized by ultraviolet light to kill any possible pathogens in the air sample. NPM-protein corona (NPC) samples were prepared as described above and dissolved in the medium by sonication. HLFs (ATCC® PCS-201-013™) were routinely grown using FGM™ fibroblast growth media (Lonza, USA) in a humidified atmosphere of 95% air and 5% CO2 at 37 °C. Cells were suspended at 2 × 106 cells/ml and mixed with the NPC solutions. NPM concentrations of NPC-HLFs were fixed at 25, 50 and 100 μg/ml, respectively. To demonstrate the bioactivity of the protein corona, transforming growth factor beta 1 (Sigma, USA), NPC-vitamin C, pure serum and pure nanoscale PM2.5 (NPM) were also mixed with HLFs following the same procedure described above. The final exposure concentrations were 10 ng/ml (TGF-β1), 0.5 mg/ml (vitamin C) and 100 μg/ml (NPM).

Manufacturer protocol Publication protocol 1 Relevant paper

D-MEM (High Glucose) with L-Glutamine and Phenol Red

Fujifilm Wako Chemicals Europe Gmbh

Upstream tips	Protocol tips	Downstream tips
	Cells were maintained in culture medium, DMEM (Wako, Tokyo, Japan) that was supplemented with 10% heat-inactivated fetal bovine serum (Thermo Fisher Scientific, Waltham, MA, USA), and 0.1 mg/ml streptomycin and 100 U/ml penicillin (Meiji Seika, Tokyo, Japan) at 37°C in a humidified atmosphere (5% CO2). the human lung adenocarcinoma epithelial cell line, A549 (European Collection of Cell Cultures, Salisbury, United Kingdom) was used to measure TGF-β1–induced decreases in E-cadherin levels, a marker of epithelial cells, in the absence or presence of a designated agent.

Protocol tips
Cells were maintained in culture medium, DMEM (Wako, Tokyo, Japan) that was supplemented with 10% heat-inactivated fetal bovine serum (Thermo Fisher Scientific, Waltham, MA, USA), and 0.1 mg/ml streptomycin and 100 U/ml penicillin (Meiji Seika, Tokyo, Japan) at 37°C in a humidified atmosphere (5% CO2). the human lung adenocarcinoma epithelial cell line, A549 (European Collection of Cell Cultures, Salisbury, United Kingdom) was used to measure TGF-β1–induced decreases in E-cadherin levels, a marker of epithelial cells, in the absence or presence of a designated agent.

Protocol tips

Cells were maintained in culture medium, DMEM (Wako, Tokyo, Japan) that was supplemented with 10% heat-inactivated fetal bovine serum (Thermo Fisher Scientific, Waltham, MA, USA), and 0.1 mg/ml streptomycin and 100 U/ml penicillin (Meiji Seika, Tokyo, Japan) at 37°C in a humidified atmosphere (5% CO2).
the human lung adenocarcinoma epithelial cell line, A549 (European Collection of Cell Cultures, Salisbury, United Kingdom) was used to measure TGF-β1–induced decreases in E-cadherin levels, a marker of epithelial cells, in the absence or presence of a designated agent.

Manufacturer protocol Publication protocol 1 Relevant paper

DMEM/F-12

Thermo Fisher Scientific

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.

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.

Manufacturer protocol Publication protocol 1 Relevant paper

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