Easy and Rapid Differentiation of Embryonic Stem Cells into Functional Motoneurons Using Sonic Hedgehog-Producing Cells

摘要: Directing embryonic stem (ES) cells to differentiate into functional motoneurons has proven be a strong technique for studying neuronal development as well being potential source of tissue cell replacement therapies involving spinal cord disorders. Unfortunately, one the mitogenic factors (i.e., sonic hedgehog agonist) used directed differentiation is not readily available, and thus this been widely accessible. Here, we present novel simple method derive from ES using attainable reagents. were derived mouse in which enhanced green fluorescent protein (eGFP) was linked motoneuron specific promoter. The plated onto monolayer 293 EcR-Shh that carry an integrated construct expression (Shh) under ecdysone-inducible control. To initiate differentiation, EcR-Shh:ES cocultures treated with ponasterone A (PA) retinoic acid 5 days. PA induces ecdysone, drives Shh expression. assess putative cell-derived studied immunocytochemically cultured on chick myotubes analysis. We found differentiated eGFP+ expressed transcription typical motoneurons. Furthermore, capable forming connections muscle fibers vitro. Finally, when transplanted developing cord, migrated ventral horn projected axons appropriate targets. In summary, treatment paradigm produces can both developmental preclinical studies. Disclosure conflicts interest at end article.