ดูโค้ด

Mouse embryonic stem (ES) cells have been served as potential model for investigation of underlying
mechanisms at cellular and molecular levels of neurological disorders. The improvement of motor neuron
differentiation is prospected to gain more understanding aimed at overcoming several incurable motor neuron
diseases. In this study, we examined the effects of selective inhibitor of TGF-β type I receptor on the efficacy of
neuronal differentiation of mouse embryonic stem cells toward motor neuron. Pluripotent ES cells were induced to
form as EB in suspension medium supplemented with retinoic acid using -4/+4 protocol. Thereafter, 8-day old EBs
were further induced to differentiate into motor neurons on monolayer culture. Our result demonstrated that the
aggregated ES cells differentiated into neuronal progenitor cells and neurons as examined with Pax-6 and Tuj-1.
Quantitative RT-PCR analysis revealed that treatment of the EBs with selective TGF-β1 inhibitor up-regulated the
motor neuron progenitor (Olig2) at higher levels than that obtained from the control (4.20±0.20 vs. 0.73±0.09, p <
0.01). In contrast, mRNA expression levels of motor neuron (Hoxc8) of the control group were significantly higher
than the TGF-β1 inhibitor treated group (14.73±2.6 vs. 2.37±0.42, p < 0.01). Immunocytochemistry demonstrated that
the differentiated cells expressed a neuronal marker (Tuj-1), motor neuron progenitor marker (Olig2) and developing
motor neuron progenitor (Isl-1), all of which are essential for generation of spinal motor neurons during neural tube
formation. Furthermore, a small proportion of differentiated cells were also positive for choline acetyltransferase
(ChAT) a marker for functional motor neurons. We concluded that modifying TGF-β signaling affected the generation
and differentiation fate of motor neuron progenitor cells.