To avoid any bias due to culture conditions, all cell lines were cultured in RPMI1640 medium with stable glutamine (Biochrom, Berlin, Germany), supplemented with 10% FCS tested to be dox-free (Sigma-Aldrich, Taufkirchen, Germany), and penicillin and streptomycin (final concentrations 100 units/mL and 100μg/mL, respectively; Merck, Darmstadt, Germany) in tissue culture flasks and plates (TPP, Trasadingen, Switzerland). To improve cell attachment for CHLA-10, CHLA-25, EW-3, EW-24, MIC, and TC-106, culture dishes were coated with 2% gelatine (Sigma-Aldrich) for cell expansion, and collagen type I solution from bovine skin (Sigma-Aldrich) in experimental assays. Cells were incubated at 37°C and 5% CO2 in a fully humidified environment. Cells were subcultured in ratios 1:2 to 1:8 after detachment with trypsin/EDTA (Biochrom), and spinning down the detached cells. Mycoplasma contamination was ruled out regularly by nested PCR with cell supernatant of each experiment.
Collagen type 1 solution from bovine skin
Manufactured by Merck Group
Collagen type I solution from bovine skin is a highly purified, sterile-filtered liquid containing type I collagen. It is extracted and purified from bovine skin. This product is suitable for cell culture applications and various research purposes.
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2 protocols using collagen type 1 solution from bovine skin
1
Standard Cell Culture Conditions
2
Standard Cell Culture Conditions
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To avoid any bias due to culture conditions, all cell lines were cultured in RPMI1640 medium with stable glutamine (Biochrom, Berlin, Germany), supplemented with 10% FCS tested to be dox-free (Sigma-Aldrich, Taufkirchen, Germany), and penicillin and streptomycin (final concentrations 100 units/mL and 100μg/mL, respectively; Merck, Darmstadt, Germany) in tissue culture flasks and plates (TPP, Trasadingen, Switzerland). To improve cell attachment for CHLA-10, CHLA-25, EW-3, EW-24, MIC, and TC-106, culture dishes were coated with 2% gelatine (Sigma-Aldrich) for cell expansion, and collagen type I solution from bovine skin (Sigma-Aldrich) in experimental assays. Cells were incubated at 37°C and 5% CO2 in a fully humidified environment. Cells were subcultured in ratios 1:2 to 1:8 after detachment with trypsin/EDTA (Biochrom), and spinning down the detached cells. Mycoplasma contamination was ruled out regularly by nested PCR with cell supernatant of each experiment.
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