Stem cells purified from human induced pluripotent stem cell-derived neural crest-like cells promote peripheral nerve regeneration

Hiroo Kimura,Taneaki Nakagawa,Tsuyoshi Amemiya,Kazuki Sato,Hideyuki Okano,Morio Matsumoto,Narihito Nagoshi,Shinsuke Shibata,Masaya Nakamura,Takehito Ouchi

doi:10.1038/s41598-018-27952-7

Hiroo Kimura, Taneaki Nakagawa + Show 8 more

Open Access

https://doi.org/10.1038/s41598-018-27952-7

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Journal: Scientific Reports	Publication Date: Jul 3, 2018
Citations: 39	License type: open-access

Affiliation: Keio University

Abstract

Strategies for therapeutic cell transplantation have been assessed for use in the treatment of massive peripheral nerve defects. To support safe and efficient cell transplantation, we have focused on the purification of cells using cell surface markers. Our group previously reported low-affinity nerve growth factor receptor (LNGFR)- and thymocyte antigen-1 (THY-1)-positive neural crest-like cells (LT-NCLCs), generated from human induced pluripotent stem cells (hiPSCs). In the present study, we investigated the efficacy of transplantation of hiPSC-derived LT-NCLCs in a murine massive peripheral nerve defect model. Animals with a sciatic nerve defect were treated with a bridging silicone tube prefilled with LT-NCLCs or medium in the transplantation (TP) and negative control (NC) groups, respectively. The grafted LT-NCLCs survived and enhanced myelination and angiogenesis, as compared to the NC group. Behavioral analysis indicated that motor functional recovery in the TP group was superior to that in the NC group, and similar to that in the autograft (Auto) group. LT-NCLCs promoted axonal regrowth and remyelination by Schwann cells. Transplantation of LT-NCLCs is a promising approach for nerve regeneration treatment of massive peripheral nerve defects.

Highlights

Tensionless nerve repair is an important advance in the surgical treatment of peripheral nerve injuries[1]
Expression level of Neural crest cells (NCCs) markers (CD49d and CD57) and Mesenchymal stem cells (MSCs) markers (CD29, CD73, CD105, CD140a, and CD146) in these low-affinity nerve growth factor receptor (LNGFR)/thymocyte antigen-1 (THY-1) single-positive and double-negative fractions were described in Table 1 and Supplementary Fig. 2a–c. These results demonstrated that the single positive fraction and double negative fraction showed the lower levels of expression for each NCC/MSC markers
The LT-NCLC-derived cells were positive for the neural markers Krox[20] and OCT6, which are markers of immature Schwann cells (Fig. 1e middle panels). They were not positive for the myelinating mature Schwann cell markers, MBP and P0 (Fig. 1e lower panels). These findings indicate that the neural crest stem/progenitors or immature Schwann cells were developed by neural crest induction

Summary

Introduction

Tensionless nerve repair is an important advance in the surgical treatment of peripheral nerve injuries[1]. Schwann cells with artificial nerve conduits improve peripheral nerve regeneration[16,17,18,19], the clinical use of such cells is limited by their source, purity, and immunologic rejection, and by potential ethical issues due to their autologous origin. We conducted the present study to identify better candidate donor cells for the treatment of massive peripheral nerve defects. We hypothesized that stem cells with the characteristics of both NCCs and MSCs might contribute to the functional recovery of massive peripheral nerve defects. Our group established a procedure for purifying a large number of LNGFR and THY-1 double-positive neural crest-like cells, designated as LT-NCLCs, from hiPSCs. The LT-NCLCs demonstrated a similar ability to NCCs and MSCs with regard to developing into Schwann-lineage cells[37]. The purpose of the present study was to assess the efficacy of LT-NCLCs derived from hiPSCs for peripheral nerve regeneration and functional recovery

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