60% confluent human T98G glioma cancer cells were split into suspension over agarose to form spheroids and grown for an additional 4 days in suspension over agarose prepared with 10% FBS and 5% non essential amino acids supplemented EMEM. Following growth, media was removed and the spheroids were stored on the surface of the agarose in vacuum sealed flasks for a 2 week interval at room temperature in the dark. The samples were compared against monolayer (at 60% confluence)samples. All samples were conducted intriplicate. Samples from the two week arrest were removed from storage and supplemented with EMEM (Invitrogen) containing 15% FBS (spheroid recovery sample). These cells were then allowed to grow out as adherent monolayers for 7 days (monolayer recovery sample).
