Notch-transfected mesenchymal stromal cells (SB623) (SanBio)

Human adult bone marrow aspirates were plated in growth medium: αMEM supplemented with fetal bovine serum (FBS; 10%) , L-glutamine (2 mM) and penicillin/streptomycin, for three days, to obtain the marrow stromal cell (MSC) monolayer. After two passages, a portion of the culture was cryopreserved as MSCs. The remaining cells (passage 2) were transfected with the pCMV-hNICD1-SV40- NeoR plasmid using Fugene6. After 7 days of selection with 100 μg/ml G418, the G418-resistant colonies were expanded and passed twice prior to cryopreservation as SB623 cells.