Human embryonic lung fibroblasts (MRC-5) was purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China). For maintenance, the cells (MRC-5) were maintained in the appropriate medium [Dulbeccos Modified Eagle Medium (DMEM)] containing 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 µg/mL streptomycin (Gibco) at 37 ℃ with 5% carbon dioxide (CO2). Before determining the concentration, we searched lots of literature and found many studies (29 (link)-31 (link)) finally determined 5 ng/mL TGF-β1 as the optimal concentration by setting different concentration and comparing the effect of promoting fibrosis. In the pre-experiment we also set different concentration of TGF-β1 (2, 5, 10 ng/mL), and found when the concentration of TGF-β1 is 5 ng/mL, the effect of promoting fibrosis is the most significant. Therefore, fibroblasts (MRC-5) were cultured in a medium supplemented with 5 ng/mL recombinant TGF-β1 (Peprotech, New Jersey, USA) for 0, 24, and 48 h to induce fibroblast activation.
Protocol detail
TGF-β1 Induced Fibrosis in MRC-5 Lung Fibroblasts
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Carbon dioxide
Cells
Cultured medium
Eagle
Embryonic
Fetal bovine serum
Fibroblast
Fibrosis
Human
Lung
Penicillin
Streptomycin
Tgf β1
Corresponding Organization : The First People's Hospital of Shunde
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Variable analysis
independent variables
- Concentration of TGF-β1 (2 ng/mL, 5 ng/mL, 10 ng/mL)
- Effect of promoting fibrosis
- Culture of human embryonic lung fibroblasts (MRC-5) in Dulbecco's Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 µg/mL streptomycin
- Incubation at 37 °C with 5% carbon dioxide (CO2)
- Not explicitly mentioned
- Not explicitly mentioned
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