The role of paraxial mesoderm or the somites in decision of ectoderm to acquire the neuroepithelial fate and its subsequent diversification to functional neural subtypes especially in mammalians is obscure. Here we report, for the first time, the influence of the co-culture of alginate bead-encapsulated somites isolated from chick embryos on neural differentiation in mouse embryonic stem cells-derived embryoid bodies (EBs). Using a combination of morphology, immunofluorescence, flow cytometry, semiquantitative, and conventional RT-PCR techniques, we show that the somites induce rosette structures and weakly enhance neural differentiation and neural markers in a dose-dependent manner in comparison to the control group, but its effect is lower than retinoic acid treatment of EBs. By liquid chromatography-mass spectrometry, it was revealed that somitic cells synthesized and released retinoids into the medium. However, no effective influence of somitic co-culture on rostrocaudal or dorsoventral patterning is seen. Therefore, like amphibians, somites have a modest neural-inducing activity in amniotes.
The role of somites and notochords in neuroectoderm differentiation from the embryonic ectoderm and its subsequent patterning into regional compartments along rostrocaudal and dorso-ventral axes, especially in humans, remains elusive. Here, we demonstrate the co-culture effect of somites and notochords isolated from chicken embryos on the neuronal differentiation and regional identity of an adherent culture of human embryonic stem cells (hESCs). Notochord increased the efficiency and speed of neuronal induction, whereas somites had a weak neuronal inducing effect on hESCs. However, a synergistic effect was not observed when notochords and somites were used together. Moreover, in somite and notochord co-culture groups, hESCs-derived neuronal cells expressed HOXB4, OTX2, IRX3 and PAX6, indicative of dorsal hindbrain and ventral anterior identities, respectively. Our results reveal the influence of embryonic notochord and somite co-culture in providing neuronal induction as well as rostrocaudal and dorso-ventral regional identity of hESCs-derived neuronal cells. This study provides a model through which in vivo neuronal induction events may be imitated.
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