The induced pluripotent stem cell (iPSc) offers great potential for cell-based therapy in regenerative medicine. We previously developed tissue-engineered bioabsorbable nerve conduits coated with iPSc-derived neurospheres for use in peripheral nerve repair. Here, we examine the long-term efficacy and safety of using nerve conduits with iPSc technology for peripheral nerve repair in mice. The nerve conduit consisted of an outer layer of a poly L-lactide mesh and an inner layer of porous sponge composed of 50% L-lactide and 50% ε-caprolactone. Secondary neurospheres were derived from mouse iPScs, resuspended and cultured within the conduit for 14 days. Conduits were implanted within surgically administered 5-mm defects in the left sciatic nerve of mice (iPSc group; n = 14). The defects in the control group (n = 13) were reconstructed using the nerve conduit alone. At 4, 8, 12, 24 and 48 weeks postsurgery, motor and sensory functional recovery in the iPSc group had improved significantly more than in the control group. At 24 and 48 weeks, histological analysis revealed axonal regeneration in the nerve conduits of both groups. However, axonal regeneration and myelination were significantly enhanced in the iPSc group. No teratomas were identified in the iPSc group at any time point. Therefore, we here demonstrate that bioabsorbable nerve conduits coated with iPSc-derived neurospheres promote enhanced regeneration of peripheral nerves and functional recovery without teratoma formation in the long term. This combination of iPSc technology and bioabsorbable nerve conduits has the potential to be a safe future tool for the treatment of peripheral nerve defects.

1.
Agnew, S.P., G.A. Dumanian (2010) Technical use of synthetic conduits for nerve repair. J Hand Surg Am 35: 838-841.
2.
Belkas, J.S., M.S. Shoichet, R. Midha (2004) Peripheral nerve regeneration through guidance tubes. Neurol Res 26: 151-160.
3.
Bertleff, M.J., M.F. Meek, J.P. Nicolai (2005) A prospective clinical evaluation of biodegradable neurolac nerve guides for sensory nerve repair in the hand. J Hand Surg Am 30: 513-518.
4.
Brockes, J.P., K.L. Fields, M.C. Raff (1979) Studies on cultured rat Schwann cells. I. Establishment of purified populations from cultures of peripheral nerve. Brain Res 165: 105-118.
5.
Carriel, V., M. Alaminos, I. Garzón, A. Campos, M. Cornelissen (2014) Tissue engineering of the peripheral nervous system. Expert Rev Neurother 14: 301-318.
6.
Chen, C.J., Y.C. Ou, S.L. Liao, W.Y. Chen, S.Y. Chen, C.W. Wu, C.C. Wang, W.Y. Wang, Y.S. Huang, S.H. Hsu (2007) Transplantation of bone marrow stromal cells for peripheral nerve repair. Exp Neurol 204: 443-453.
7.
Daly, W., L. Yao, D. Zeugolis, A. Windebank, A. Pandit (2012) A biomaterials approach to peripheral nerve regeneration: bridging the peripheral nerve gap and enhancing functional recovery. J R Soc Interface 9: 202-221.
8.
Derby, A., V.W. Engleman, G.E. Frierdich, G. Neises, S.R. Rapp, D.G. Roufa (1993) Nerve growth factor facilitates regeneration across nerve gaps: morphological and behavioral studies in rat sciatic nerve. Exp Neurol 119: 176-191.
9.
George, L.T., T.M. Myckatyn, J.N. Jensen, D.A. Hunter, S.E. Mackinnon (2003) Functional recovery and histomorphometric assessment following tibial nerve injury in the mouse. J Reconstr Microsurg 19: 41-48.
10.
Griffin, J.W., M.V. Hogan, A.B. Chhabra, D.N. Deal (2013) Peripheral nerve repair and reconstruction. J Bone Joint Surg Am 95: 2144-2151.
11.
Guénard, V., N. Kleitman, T.K. Morrissey, R.P. Bunge, P. Aebischer (1992) Syngeneic Schwann cells derived from adult nerves seeded in semipermeable guidance channels enhance peripheral nerve regeneration. J Neurosci 12: 3310-3320.
12.
Hadlock, T., C. Sundback, D. Hunter, M. Cheney, J.P. Vacanti (2000) A polymer foam conduit seeded with Schwann cells promotes guided peripheral nerve regeneration. Tissue Eng 6: 119-127.
13.
Hsu, Y.C., S.L. Chen, D.Y. Wang, I.M. Chiu (2013) Stem cell-based therapy in neural repair. Biomed J 36: 98-105.
14.
Hu, N., H. Wu, C. Xue, Y. Gong, J. Wu, Z. Xiao, Y. Yang, F. Ding, X. Gu (2013) Long-term outcome of the repair of 50 mm long median nerve defects in rhesus monkeys with marrow mesenchymal stem cells-containing, chitosan-based tissue engineered nerve grafts. Biomaterials 34: 100-111.
15.
Ikeda, M., T. Uemura, K. Takamatsu, M. Okada, K. Kazuki, Y. Tabata, Y. Ikada, H. Nakamura (2014) Acceleration of peripheral nerve regeneration using nerve conduits in combination with induced pluripotent stem cell technology and a basic fibroblast growth factor drug delivery system. J Biomed Mater Res A 102: 1370-1378.
16.
Inoue, H., S. Yamanaka (2011) The use of induced pluripotent stem cells in drug development. Clin Pharmacol Ther 89: 655-661.
17.
Jiang, X., S.H. Lim, H.Q. Mao, S.Y. Chew (2010) Current applications and future perspectives of artificial nerve conduits. Exp Neurol 223: 86-101.
18.
Johnson, P.J., M.D. Wood, A.M. Moore, S.E. Mackinnon (2013) Tissue engineered constructs for peripheral nerve surgery. Eur Surg 45: 270-276.
19.
Jungnickel, J., K. Haase, J. Konitzer, M. Timmer, C. Grothe (2006) Faster nerve regeneration after sciatic nerve injury in mice over-expressing basic fibroblast growth factor. J Neurobiol 66: 940-948.
20.
Kehoe, S., X.F. Zhang, D. Boyd (2012) FDA approved guidance conduits and wraps for peripheral nerve injury: a review of materials and efficacy. Injury 43: 553-572.
21.
Kobayashi, Y., Y. Okada, G. Itakura, H. Iwai, S. Nishimura, A. Yasuda, S. Nori, K. Hikishima, T. Konomi, K. Fujiyoshi, O. Tsuji, Y. Toyama, S. Yamanaka, M. Nakamura, H. Okano (2012) Pre-evaluated safe human iPSC-derived neural stem cells promote functional recovery after spinal cord injury in common marmoset without tumorigenicity. PLoS One 7: e52787.
22.
Koshimune, M., K. Takamatsu, H. Nakatsuka, K. Inui, Y. Yamano, Y. Ikada (2003) Creating bioabsorbable Schwann cell coated conduits through tissue engineering. Biomed Mater Eng 13: 223-229.
23.
Li, R., Z. Liu, Y. Pan, L. Chen, Z. Zhang, L. Lu (2014) Peripheral nerve injuries treatment: a systematic review. Cell Biochem Biophys 68: 449-454.
24.
Marchesi, C., M. Pluderi, F. Colleoni, M. Belicchi, M. Meregalli, A. Farini, D. Parolini, L. Draghi, M.E. Fruguglietti, M. Gavina, L. Porretti, A. Cattaneo, M. Battistelli, A. Prelle, M. Moggio, S. Borsa, L. Bello, D. Spagnoli, S.M. Gaini, M.C. Tanzi, N. Bresolin, N. Grimoldi, Y. Torrente (2007) Skin-derived stem cells transplanted into resorbable guides provide functional nerve regeneration after sciatic nerve resection. Glia 55: 425-438.
25.
Miura, K., Y. Okada, T. Aoi, A. Okada, K. Takahashi, K. Okita, M. Nakagawa, M. Koyanagi, K. Tanabe, M. Ohnuki, D. Ogawa, E. Ikeda, H. Okano, S. Yamanaka (2009) Variation in the safety of induced pluripotent stem cell lines. Nat Biotechnol 27: 743-745.
26.
Moore, A.M., R. Kasukurthi, C.K. Magill, H.F. Farhadi, G.H. Borschel, S.E. Mackinnon (2009) Limitations of conduits in peripheral nerve repairs. Hand 4: 180-186.
27.
Mosahebi, A., P. Fuller, M. Wiberg, G. Terenghi (2002) Effect of allogeneic Schwann cell transplantation on peripheral nerve regeneration. Exp Neurol 173: 213-223.
28.
Nakagawa, M., M. Koyanagi, K. Tanabe, K. Takahashi, T. Ichisaka, T. Aoi, K. Okita, Y. Mochiduki, N. Takizawa, S. Yamanaka (2008) Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts. Nat Biotechnol 26: 101-106.
29.
Nakamura, M., H. Okano (2013) Cell transplantation therapies for spinal cord injury focusing on induced pluripotent stem cells. Cell Res 23: 70-80.
30.
Nelson, T.J., A. Martinez-Fernandez, S. Yamada, C. Perez-Terzic, Y. Ikeda, A. Terzic (2009) Repair of acute myocardial infarction by human stemness factors induced pluripotent stem cells. Circulation 120: 408-416.
31.
Nishimura, K., J. Takahashi (2013) Therapeutic application of stem cell technology toward the treatment of Parkinson's disease. Biol Pharm Bull 36: 171-175.
32.
Nori, S., Y. Okada, A. Yasuda, O. Tsuji, Y. Takahashi, Y. Kobayashi, K. Fujiyoshi, M. Koike, Y. Uchiyama, E. Ikeda, Y. Toyama, S. Yamanaka, M. Nakamura, H. Okano (2011) Grafted human-induced pluripotent stem-cell-derived neurospheres promote motor functional recovery after spinal cord injury in mice. Proc Natl Acad Sci USA 108: 16825-16830.
33.
Okada, Y., A. Matsumoto, T. Shimazaki, R. Enoki, A. Koizumi, S. Ishii, Y. Itoyama, G. Sobue, H. Okano (2008) Spatiotemporal recapitulation of central nervous system development by murine embryonic stem cell-derived neural stem/progenitor cells. Stem Cells 26: 3086-3098.
34.
Okano, H., M. Nakamura, K. Yoshida, Y. Okada, O. Tsuji, S. Nori, E. Ikeda, S. Yamanaka, K. Miura (2013) Steps toward safe cell therapy using induced pluripotent stem cells. Circ Res 112: 523-533.
35.
Okita, K., T. Ichisaka, S. Yamanaka (2007) Generation of germline-competent induced pluripotent stem cells. Nature 448: 313-317.
36.
Okita, K., M. Nakagawa, H. Hyenjong, T. Ichisaka, S. Yamanaka (2008) Generation of mouse induced pluripotent stem cells without viral vectors. Science 322: 949-953.
37.
Okita, K., S. Yamanaka (2011) Induced pluripotent stem cells: opportunities and challenges. Philos Trans R Soc Lond B Biol Sci 366: 2198-2207.
38.
Pabari, A., S.Y. Yang, A.M. Seifalian, A. Mosahebi (2010) Modern surgical management of peripheral nerve gap. J Plast Reconstr Aesthet Surg 63: 1941-1948.
39.
Pereira Lopes, F.R., L. Camargo de Moura Campos, J. Dias Corrêa, A. Balduino, S. Lora, F. Langone, R. Borojevic, A.M. Blanco Martinez (2006) Bone marrow stromal cells and resorbable collagen guidance tubes enhance sciatic nerve regeneration in mice. Exp Neurol 198: 457-468.
40.
Prockop, D.J. (2010) Defining the probability that a cell therapy will produce a malignancy. Mol Ther 18: 1249-1250.
41.
Suemori, H., K. Yasuchika, K. Hasegawa, T. Fujioka, N. Tsuneyoshi, N. Nakatsuji (2006) Efficient establishment of human embryonic stem cell lines and long-term maintenance with stable karyotype by enzymatic bulk passage. Biochem Biophys Res Commun 345: 926-932.
42.
Takahashi, K., S. Yamanaka (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126: 663-676.
43.
Thomson, J.A., V.S. Marshall (1998) Primate embryonic stem cells. Curr Top Dev Biol 38: 133-165.
44.
Thomson, J.A., J.S. Odorico (2000) Human embryonic stem cell and embryonic germ cell lines. Trends Biotechnol 18: 53-57.
45.
Tohill, M., C. Mantovani, M. Wiberg, G. Terenghi (2004) Rat bone marrow mesenchymal stem cells express glial markers and stimulate nerve regeneration. Neurosci Lett 362: 200-203.
46.
Tsuji, O., K. Miura, Y. Okada, K. Fujiyoshi, M. Mukaino, N. Nagoshi, K. Kitamura, G. Kumagai, M. Nishino, S. Tomisato, H. Higashi, T. Nagai, H. Katoh, K. Kohda, Y. Matsuzaki, M. Yuzaki, E. Ikeda, Y. Toyama, M. Nakamura, S. Yamanaka, H. Okano (2010) Therapeutic potential of appropriately evaluated safe-induced pluripotent stem cells for spinal cord injury. Proc Natl Acad Sci USA 107: 12704-12709.
47.
Uemura, T., K. Takamatsu, M. Ikeda, M. Okada, K. Kazuki, Y. Ikada, H. Nakamura (2011) A tissue-engineered bioabsorbable nerve conduit created by three-dimensional culture of induced pluripotent stem cell-derived neurospheres. Biomed Mater Eng 21: 333-339.
48.
Uemura, T., K. Takamatsu, M. Ikeda, M. Okada, K. Kazuki, Y. Ikada, H. Nakamura (2012) Transplantation of induced pluripotent stem cell-derived neurospheres for peripheral nerve repair. Biochem Biophys Res Commun 419: 130-135.
49.
Wangensteen, K.J., L.K. Kalliainen (2010) Collagen tube conduits in peripheral nerve repair: a retrospective analysis. Hand 5: 273-277.
50.
Weber, R.A., W.C. Breidenbach, R.E. Brown, M.E. Jabaley, D.P. Mass (2000) A randomized prospective study of polyglycolic acid conduits for digital nerve reconstruction in humans. Plast Reconstr Surg 106: 1036-1048.
51.
Wernig, M., A. Meissner, J.P. Cassady, R. Jaenisch (2008) c-Myc is dispensable for direct reprogramming of mouse fibroblasts. Cell Stem Cell 2: 10-12.
52.
Xu, L., S. Zhou, G.Y. Feng, L.P. Zhang, D.M. Zhao, Y. Sun, Q. Liu, F. Huang (2012) Neural stem cells enhance nerve regeneration after sciatic nerve injury in rats. Mol Neurobiol 46: 265-274.
53.
Xu, W., C.S. Cox, Y. Li (2011) Induced pluripotent stem cells for peripheral nerve regeneration. J Stem Cells 6: 39-49.
54.
Yamashita, T., H. Kawai, F. Tian, Y. Ohta, K. Abe (2011) Tumorigenic development of induced pluripotent stem cells in ischemic mouse brain. Cell Transplant 20: 883-891.
55.
Yoshida, Y., S. Yamanaka (2010) Recent stem cell advances: induced pluripotent stem cells for disease modeling and stem cell-based regeneration. Circulation 122: 80-87.
56.
Zhang, Y., D. Wang, M. Chen, B. Yang, F. Zhang, K. Cao (2011) Intramyocardial transplantation of undifferentiated rat induced pluripotent stem cells causes tumorigenesis in the heart. PLoS One 6: e19012.
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.