Patent ID: 8058063

Claim:
A method of providing cholinergic neurons to a subject affected by a condition characterized by the loss of cholinergic neurons, the method comprising: expanding human neural stem cells and/or progeny thereof by suspension culture as neurospheres in a serum-free mixture comprising epidermal growth factor, basic fibroblast growth factor and leukemia inhibitory factor (LIF); adhesively culturing the expanded human neural stem cells and/or progeny thereof in vitro with a serum-free mixture comprising basic fibroblast growth factor, mammalian Sonic Hedgehog protein or an N-terminus-containing fragment thereof, laminin and heparin; and implanting the adhesively cultured human neural stem cells and/or progeny thereof into the spinal cord or brain of the subject, wherein the implanted adhesively cultured human neural stem cells and/or progeny thereof differentiate into cholinergic neurons and wherein the subject is affected with ALS or a spinal cord injury.