In this study, hair follicle neural crest stem cells (hfNCSC) in rats were induced into neurons by sonic hedgehog/retino acid (Shh/RA) and Schwann cells by neuregulin-1 (NRG-1). Then suspension of these neurons and Schwann cells were mixed with a specific ratio and implanted into a acellular nerve graft to construct a tissue engineering nerve graft. Examination was performed on the nerve graft in vitro and in vivo. This manuscript was divided into five sections as follows:isolation, culture and induction as well as identification of hfNCSC; preparation of acellular nerve graft in Beagles; construction of nerve graft in vitro, culture of nerve graft in vitro, transplantation and examination of tissue engineering nerve graft in vivo. This study validated that neurons derived from stem cells can survive and keep neuron characteristics in peripheral nerves. Formation of synapse and nerve circuit between implanted neurons were observed in peripheral nerves.
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