Motor neurons suspension-based differentiation of human embryonic stem cells

In this differentiation protocol, human embryonic stem cells are differentiated into motor neuron progenitor cells via a suspension-based procedure and the formation of neurospheres. The differentiated cells express specific motor neurons markers, such as MNX1 (HB9), Isl1, beta-III-tubulin, and ChAT (in long-term cultures). In addition, a conditioned medium, produced from the motor neurons cultures, contains neurotrophic factors such as nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), vascular endothelial growth factor (VEGF), neurotrophin 3 (NT-3), and neurotrophin 4 (NT-4). To investigate their functional properties, the differentiated cells were transplanted into rodent models of spinal cord injury (SCI), spinal muscular atrophy (SMA), and amyotrophic lateral sclerosis (ALS).