1C115-HT cell culture was performed as described previously(82 (link)). Briefly, undifferentiated 1C11 cells were kept on 100 mm plates (Sarstedt) in DMEM Glutamax with 10% fetal bovine serum, 1% non-essential amino acids, 1% penicillin/streptomycin, and 1% L-glutamine (all media components by Life Technologies) at 37°C and 5% CO2. For differentiation to serotonin neuron-like cells (1C115-HT) 10.000 cells were transferred to 8-well imaging slides (Ibidi). Then, culture medium was supplemented with 1 mM dibutyryl cAMP and 0.05% cyclohexanecarboxylic acid for 4 days (media supplements by Sigma Aldrich). Treatment conditions: For determination of SERT cell surface density, medium was supplemented with 15 mM KCl and/or 1 μM escitalopram. For ASP+ live cell imaging, cells were transferred to FSCV buffer containing 50 μM ASP and/or 1 μM escitalopram and/or serotonin (0.1 or 1 μM). Electrical stimulation was applied after perfusing the imaging slides with ASP+-containing FSCV buffer. Dye-free buffer was applied 1 min after electrical stimulation before image acquisition.
8 well imaging slides
Manufactured by Ibidi
The 8-well imaging slides are a laboratory product designed for cell-based studies and imaging applications. Each slide features eight individual wells, allowing for the simultaneous cultivation and observation of multiple cell samples or experimental conditions.
Automatically generated - may contain errors
2 protocols using 8 well imaging slides
1
Differentiation and Characterization of 1C11 Serotonin Neurons
2
1C11 Serotonin Neuron Differentiation
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1C11 5-HT cell culture was performed as described previously (82) . Briefly, undifferentiated 1C11 cells were kept on 100 mm plates (Sarstedt) in DMEM Glutamax with 10% fetal bovine serum, 1% non-essential amino acids, 1% penicillin/streptomycin, and 1% L-glutamine (all media components by Life Technologies) at 37 °C and 5% CO 2 . For differentiation to serotonin neuron-like cells (1C11 5-HT ) 10.000 cells were transferred to 8-well imaging slides (Ibidi). Then, culture medium was supplemented with 1 mM dibutyryl cAMP and 0.05% cyclohexanecarboxylic acid for 4 days (media supplements by Sigma Aldrich). Treatment conditions: For determination of SERT cell surface density, medium was supplemented with 15 mM KCl and/or 1 μM escitalopram. For ASP + live cell imaging, cells were transferred to FSCV buffer containing 50 μM ASP and/or 1 μM escitalopram and/or serotonin (0.1 or 1 μM). Electrical stimulation was applied after perfusing the imaging slides with ASP + -containing FSCV buffer. Dye-free buffer was applied 1 min after electrical stimulation before image acquisition.
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