Motoneurons derived from embryonic stem cells express transcription factors and develop phenotypes characteristic of medial motor column neurons

摘要: Embryonic stem (ES) cells differentiate into functional motoneurons when treated with a sonic hedgehog (Shh) agonist and retinoic acid (RA). Whether ES can be directed to specific subtypes of is unknown. We embryoid bodies generated from HBG3 Shh RA for 5 d in culture induce motoneuron differentiation. Enhanced green fluorescent protein (eGFP) expression was used identify putative motoneurons, because eGFP expressed under the control Hb9 promoter cells. found that 96 +/- 0.7% differentiated eGFP+ Lhx3, homeobox gene by postmitotic medial motor column (MMCm), were plated on neurite-promoting substrate d. When transplanted stage 17 chick neural tubes, migrated MMCm, projected axons appropriate target MMCm (i.e., epaxial muscles), contained synaptic vesicles within intramuscular axonal branches. In ovo vitro studies indicated chemotropic factors emanating muscle and/or surrounding mesenchyme likely guide Lhx3+ their correct target. Finally, whole-cell patch-clamp recordings cell-derived demonstrated they received input, elicited repetitive trains action potentials, developed passive membrane properties similar host motoneurons. These results indicate form neurons phenotypic properties.