Direct conversion of adult human skin fibroblasts into functional Schwann cells that achieve robust recovery of the severed peripheral nerve in rats

• Published in: Glia 2019-05, Vol.67 (5), p.950-966
• Main Authors: Kitada, Masaaki, Murakami, Toru, Wakao, Shohei, Li, Gen, Dezawa, Mari
• Format: Article
• Language: English
• Subjects: axonal regeneration; Axonogenesis; Axons; Bone marrow; Central nervous system; Demyelinating diseases; Demyelination; Direct conversion; Fibroblast growth factor 2; Fibroblasts; Forskolin; Gene expression; Genomes; Growth factors; Heregulin; Medical treatment; Mesenchyme; Myelination; Neuromuscular junctions; PNS injury; Recovery; Recovery of function; Regeneration; Retinoic acid; re‐formation of neuromuscular junction; Schwann cells; Stem cells; Therapeutic applications; Transplantation; Tumorigenesis
• ISSN: 0894-1491; 1098-1136
• DOI: 10.1002/glia.23582

Direct conversion is considered a promising approach to obtain tissue‐specific cells for cell therapies; however, this strategy depends on exogenous gene expression that may cause undesired adverse effects such as tumorigenesis. By optimizing the Schwann cell induction system, which was originally developed for trans‐differentiation of bone marrow mesenchymal stem cells into Schwann cells, we established a system to directly convert adult human skin fibroblasts into cells comparable to authentic human Schwann cells without gene introduction. Serial treatments with beta‐mercaptoethanol, retinoic acid, and finally a cocktail of basic fibroblast growth factor, forskolin, platelet‐derived growth factor‐AA, and heregulin‐β1 (EGF domain) converted fibroblasts into cells expressing authentic Schwann cell markers at an efficiency of approximately 75%. Genome‐wide gene expression analysis suggested the conversion of fibroblasts into the Schwann cell‐lineage. Transplantation of induced Schwann cells into severed peripheral nerve of rats facilitated axonal regeneration and robust functional recovery in sciatic function index comparable to those of authentic human Schwann cells. The contributions of induced Schwann cells to myelination of regenerated axons and re‐formation of neuromuscular junctions were also demonstrated. Our data clearly demonstrated that cells comparable to functional Schwann cells feasible for the treatment of neural disease can be induced from adult human skin fibroblasts without gene introduction. This direct conversion system will be beneficial for clinical applications to peripheral and central nervous system injuries and demyelinating diseases. Main points Under the specific induction system without gene transfer, adult human skin fibroblasts can be directly converted into cells comparable to authentic Schwann cells, which facilitate robust nerve regeneration in the severed sciatic nerve of rats.