BMDMs were isolated from femora bones of adult mice. 5x105 cells were plated 10 cm-diameter tissue culture dish in L929-CM (20%). Cells were cultivated (DMEM, 1% P/S, 10% fetal calf serum) at 37 °C with 7.5% CO2 v/v. In vitro recombination was achieved by 4-hydroxytamoxifen (Sigma-Aldrich) administration at a final dose of 4 μM for 10 days (dissolved in EtOH) starting 24h after plating. For experimental seeding, cells were harvested following accutase (Innovative Cell Technologies Inc.) induced detachment. Cells were either seeded into cell culture grade plasticware, onto Poly-L-lysine (PLL) coated, HCl-washed 1.5# 12 mm diameter glass coverslips, or into PLL coated glass bottom chamber slides (Ibidi) for live imaging. Cells were seeded (6-well: 250k cells; 96-well: 18k cells; 12mm coverslip: 18k cells; 8-chamber slide: 18k cells) in cultivation media. For activation, cells were treated with LPS (10 ng/mL) and/or myelin (10 μg/mL) if not otherwise stated. Squalene (100 μM in HBSS) or DMHCA (10 μM in DMF) were added 24h before activation.
Pll coated glass bottom chamber slides
Manufactured by Ibidi
The PLL coated glass bottom chamber slides are a specialized laboratory equipment designed for cell culture applications. These slides feature a glass bottom with a poly-L-lysine (PLL) coating, which facilitates the adhesion and growth of cells. The PLL coating provides a suitable surface for cell attachment, enabling researchers to observe and study cell behavior in a controlled environment.
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Lab products found in correlation
2 protocols using pll coated glass bottom chamber slides
1
Isolation and In Vitro Activation of Murine BMDMs
2
Isolation and In Vitro Activation of Murine BMDMs
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BMDMs were isolated from femora bones of adult mice. 5x105 cells were plated 10 cm-diameter tissue culture dish in L929-CM (20%). Cells were cultivated (DMEM, 1% P/S, 10% fetal calf serum) at 37 °C with 7.5% CO2 v/v. In vitro recombination was achieved by 4-hydroxytamoxifen (Sigma-Aldrich) administration at a final dose of 4 μM for 10 days (dissolved in EtOH) starting 24h after plating. For experimental seeding, cells were harvested following accutase (Innovative Cell Technologies Inc.) induced detachment. Cells were either seeded into cell culture grade plasticware, onto Poly-L-lysine (PLL) coated, HCl-washed 1.5# 12 mm diameter glass coverslips, or into PLL coated glass bottom chamber slides (Ibidi) for live imaging. Cells were seeded (6-well: 250k cells; 96-well: 18k cells; 12mm coverslip: 18k cells; 8-chamber slide: 18k cells) in cultivation media. For activation, cells were treated with LPS (10 ng/mL) and/or myelin (10 μg/mL) if not otherwise stated. Squalene (100 μM in HBSS) or DMHCA (10 μM in DMF) were added 24h before activation.
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