2.2. Preparation of conditioned medium and conditioning of MSCs

Conditioned medium was obtained by culturing SH-SY5Y cells in complete DMEM-F12 medium incubated at 37 °C with 5% CO₂ in a humidified environment. The medium was changed every 3 days and once confluent the spent medium was collected and centrifuged at 500 g for 15 min. Medium was stored at 2–8 °C for up to seven days.

MSCs were conditioned by the addition of equal volumes of complete medium and spent medium obtained from SH-SY5Y cultures. These were kept incubated at 37 °C with 5% CO₂ in a humidified environment and medium was changed every 3 days. Morphological changes could be seen within 24 h. Such cells are referred to hereafter as Conditioned Cells (CCs).