Example protocol for Gelacell™ nanofiber scaffold Plate
Cell seeding and culture
1. Before cell seeding, rinse the PLGA:PCL scaffold once with 70% ethanol (see table below), followed by a rinse with PBS solution three times.
2. Pre-swell the scaffold with your preferred cell culture media (with/without serum). Add culture media (see table below) onto the scaffold, incubate for 30 mins to 1 hour, and aspirate the spent media.
3. After pre-swelling, allow the scaffold to partially dry in a laminar hood by keeping the lid open for 20 minutes. It helps in absorbing the cell seeding volume and spreading the cells uniformly.
4. Dispense your desired concentration of cell culture suspension with complete media onto the scaffold. The range of cell seeding density is given in table below as a recommendation, but the exact figure will depend on the cell type and the planned duration of cell culture.
5. Place the well in a CO2-incubator at 37°C for 30 mins to 2 hours to allow initial cell adhesion. Afterward, gently fill the well with medium (see table below) from the sides of the well without dislodging the cells that have already adhered to the scaffold. Return the scaffolds in the incubator for culturing cells.
6. It is recommended to exchange media roughly every 24 hours to 48 hours; however, this may vary depending on the cell line, media, and cell density. Aspirate the waste media from the sides of the well and carefully add fresh media (see table below) on top of the scaffold without dislodging the adhered cells. Continue media exchange throughout the cell culture period.