Culturing of Daoy cells in astrocyte conditioned media

Astrocytes are important cells within the medulloblastoma microenvironment. Therefore, we assessed how astrocyte secreted factors may alter Daoy (human MB cell line; ATCC HTB-186) cell gene expression. Astrocytes were isolated from the hippocampi of 1-2 day old Sprague Dawley rat pups and cultured in vitro. Supernatent, or astrocyte conditioned media (ACM) was collected from the astrocyte cultures every 2-3 days. This supernatent was used to culture Daoy (ATCC HTB-186) cells for 1 (P1) and 3 (P3)passages. Each passage was cultured for 48 h. For control samples, Daoy cells were cultured in the same manner but using full DMEM (DM) media. Three independant experiments were performed.