Background: Neural crest (NC) cells emerge at the interface between neural and non-neural ectoderm, and migrate extensively to form a variety of NC derivatives such as peripheral neurons, glia, melanocytes, endocrine cells, and mesenchymal precursor cells. NC cells possess various unique properties and are capable of undergoing cell fate decisions across multiple tissues and germ layers. In zebrafish and mouse, GnRH neurons are reported to arise also from NC.
Objective and hypotheses: Here we show that GnRH neuron-like cells can also be obtained from NC cells, which are derived from human pluripotent stem cells (hPSCs).
Method: We first induced NC fate from hPSCs by our recently published protocol; these hPSC-derived NC cells expressed NC-specifier genes, including MSX2, PAX3, SLUG, TWIST1, and SOX10, and were confirmed to be multipotent. We next differentiated hPSC-derived NC cells toward neuronal lineage, which resulted in upregulation of a set of neuronal genes including TUJ1, MASH1, and NGN2.
Results: Peripheral sensory neurons and sympathetic neurons were detected by immunocytochemistry. Importantly, genes related to GnRH neuron development such as EBF2, DCC, and VAX1 were increased upon neuronal induction. GnRH1 expression did not increase significantly, but GnRH1-immunopositive cells were detected among TUJ1-positive neurons. The resulting GnRH1-positive cells were co-localized with NC markers, such as SOXE, p75(NGFR) and HNK1.
Conclusion: GnRH-expressing cells can be generated from hPSC-derived NC cells. We are currently investigating the implications of these findings with Kallmann syndrome patient-derived induced pluripotent stem cells.
20 - 22 Sep 2014
European Society for Paediatric Endocrinology