Background. Autologous transplantation is an attractive approach to treat sortie neurological diseases. A major obstacle is the capacity to produce cells for transplantation at the appropriate time. We describe a cryopreservation procedure for adult human brain tissue allowing the generation of cells in vitro. Methods. Neurological resections were dissected to separate white and grey matter. Fractions were frozen in a specific cryopreservation medium containing a selected serum and stored in liquid nitrogen. Tissue was thawed, cells were mechanically dissociated, expanded ill Culture and characterized by immunochemistry. Results. Adult human brain tissue cryopreserved for LIP to two years was successfully used to generate brain cells that could be maintained in culture for LIP to 100 days. Cells expressed a variety of neuroectodermal markers including GFAP, S100beta, and neurofilament. Conclusion. A successful procedure for cryopreservation of adult human brain tissue has been established that might facilitate future autologous transplantation strategies. (C) 2003 Published by Elsevier Inc.