title: Adipose-Derived Stem Cells Promote Peripheral Nerve Regeneration In Vivo without Differentiation into Schwann-Like Lineage.
abstract: BACKGROUND
  During recent decades, multipotent stem cells were found to reside in the adipose tissue, and these adipose-derived stem cells were shown to play beneficial roles, like those of Schwann cells, in peripheral nerve regeneration. However, it has not been well established whether adipose-derived stem cells offer beneficial effects to peripheral nerve injuries in vivo as Schwann cells do. Furthermore, the in situ survival and differentiation of adipose-derived stem cells after transplantation at the injured peripheral nerve tissue remain to be fully elucidated.
  
  
  METHODS
  Adipose-derived stem cells and Schwann cells were transplanted with gelatin hydrogel tubes at the artificially blunted sciatic nerve lesion in mice. Neuroregenerative abilities of them were comparably estimated. Cre-loxP-mediated fate tracking was performed to visualize survival in vivo of transplanted adipose-derived stem cells and to investigate whether they differentiated into Schwann linage cells at the peripheral nerve injury site.
  
  
  RESULTS
  The transplantation of adipose-derived stem cells promoted regeneration of axons, formation of myelin, and restoration of denervation muscle atrophy to levels comparable to those achieved by Schwann cell transplantation. The adipose-derived stem cells survived for at least 4 weeks after transplantation without differentiating into Schwann cells.
  
  
  CONCLUSIONS
  Transplanted adipose-derived stem cells did not differentiate into Schwann cells but promoted peripheral nerve regeneration at the injured site. The neuroregenerative ability was comparable to that of Schwann cells. Adipose-derived stem cells at an undifferentiated stage may be used as an alternative cell source for autologous cell therapy for patients with peripheral nerve injury.
authors.affiliation: Kyoto and Fukushima, Japan From the Departments of Plastic and Reconstructive Surgery and Immunology, Kyoto Prefectural University of Medicine, Graduate School of Medical Sciences; the Department of Biomaterials, Field of Tissue Engineering, Institute for Frontier Medical Sciences, Kyoto University; and the Department of Basic Pathology, Fukushima Medical University School of Medicine.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Sowa
authors.forename: Tsunao
authors.initials: T
authors.lastname: Kishida
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Imura
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Numajiri
authors.forename: Kenichi
authors.initials: K
authors.lastname: Nishino
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Tabata
authors.forename: Osam
authors.initials: O
authors.lastname: Mazda
mesh_terms: D017667:Adipocytes; D000818:Animals; D002454:Cell Differentiation; D051379:Mice; D009416:Nerve Regeneration; D059348:Peripheral Nerve Injuries / Q000601:surgery*; D010525:Peripheral Nerves / Q000502:physiology*; D012583:Schwann Cells; D033581:Stem Cell Transplantation
references.citation: Mackinnon SE, Hudson AR. Clinical application of peripheral nerve transplantation. Plast Reconstr Surg. 1992;90:695–699
references.citation: Chen ZL, Yu WM, Strickland S. Peripheral regeneration. Annu Rev Neurosci. 2007;30:209–233
references.citation: Hadlock T, Sundback C, Hunter D, Cheney M, Vacanti JP. A polymer foam conduit seeded with Schwann cells promotes guided peripheral nerve regeneration. Tissue Eng. 2000;6:119–127
references.citation: Faroni A, Terenghi G, Reid AJ. Adipose-derived stem cells and nerve regeneration: Promises and pitfalls. Int Rev Neurobiol. 2013;108:121–136
references.citation: Zuk PA, Zhu M, Mizuno H, et al. Multilineage cells from human adipose tissue: Implications for cell-based therapies. Tissue Eng. 2001;7:211–228
references.citation: Fraser JK, Wulur I, Alfonso Z, Hedrick MH. Fat tissue: An underappreciated source of stem cells for biotechnology. Trends Biotechnol. 2006;24:150–154
references.citation: Gimble JM, Katz AJ, Bunnell BA. Adipose-derived stem cells for regenerative medicine. Circ Res. 2007;100:1249–1260
references.citation: Orbay H, Uysal AC, Hyakusoku H, Mizuno H. Differentiated and undifferentiated adipose-derived stem cells improve function in rats with peripheral nerve gaps. J Plast Reconstr Aesthet Surg. 2012;65:657–664
references.citation: Shen CC, Yang YC, Liu BS. Peripheral nerve repair of transplanted undifferentiated adipose tissue-derived stem cells in a biodegradable reinforced nerve conduit. J Biomed Mater Res A. 2012;100:48–63
references.citation: Zhang HT, Luo J, Sui LS, et al. Effects of differentiated versus undifferentiated adipose tissue-derived stromal cell grafts on functional recovery after spinal cord contusion. Cell Mol Neurobiol. 2009;29:1283–1292
references.citation: Qian DX, Zhang HT, Ma X, Jiang XD, Xu RX. Comparison of the efficiencies of three neural induction protocols in human adipose stromal cells. Neurochem Res. 2010;35:572–579
references.citation: Kalbermatten DF, Schaakxs D, Kingham PJ, Wiberg M. Neurotrophic activity of human adipose stem cells isolated from deep and superficial layers of abdominal fat. Cell Tissue Res. 2011;344:251–260
references.citation: Sowa Y, Imura T, Numajiri T, Nishino K, Fushiki S. Adipose-derived stem cells produce factors enhancing peripheral nerve regeneration: Influence of age and anatomic site of origin. Stem Cells Dev. 2012;21:1852–1862
references.citation: Wang H, Zhang H, Liu M, Wang N. Distal segment extracts of the degenerated rat sciatic nerve induce bone marrow stromal cells to express Schwann cell markers in vitro. Neurosci Lett. 2013;544:89–93
references.citation: Yang J, Lou Q, Huang R, Shen L, Chen Z. Dorsal root ganglion neurons induce transdifferentiation of mesenchymal stem cells along a Schwann cell lineage. Neurosci Lett. 2008;445:246–251
references.citation: Gilchrist DS, Ure J, Hook L, Medvinsky A. Labeling of hematopoietic stem and progenitor cells in novel activatable EGFP reporter mice. Genesis. 2003;36:168–176
references.citation: Dinh P, Hazel A, Palispis W, Suryadevara S, Gupta R. Functional assessment after sciatic nerve injury in a rat model. Microsurgery. 2009;29:644–649
references.citation: Herrmann JE, Imura T, Song B, et al. STAT3 is a critical regulator of astrogliosis and scar formation after spinal cord injury. J Neurosci. 2008;28:7231–7243
references.citation: Kawamoto S, Niwa H, Tashiro F, et al. A novel reporter mouse strain that expresses enhanced green fluorescent protein upon Cre-mediated recombination. FEBS Lett. 2000;470:263–268
references.citation: Muzumdar MD, Tasic B, Miyamichi K, Li L, Luo L. A global double-fluorescent Cre reporter mouse. Genesis. 2007;45:593–605
references.citation: Peng LH, Mao ZY, Qi XT, et al. Transplantation of bone-marrow-derived mesenchymal and epidermal stem cells contribute to wound healing with different regenerative features. Cell Tissue Res. 2013;352:573–583
references.citation: Yamamoto M, Ikada Y, Tabata Y. Controlled release of growth factors based on biodegradation of gelatin hydrogel. J Biomater Sci Polym Ed. 2001;12:77–88
references.citation: Taketo M, Schroeder AC, Mobraaten LE, et al. FVB/N: An inbred mouse strain preferable for transgenic analyses. Proc Natl Acad Sci USA. 1991;88:2065–2069
references.citation: Clavijo-Alvarez JA, Nguyen VT, Santiago LY, Doctor JS, Lee WP, Marra KG. Comparison of biodegradable conduits within aged rat sciatic nerve defects. Plast Reconstr Surg. 2007;119:1839–1851
references.citation: Yuan Y, Shen H, Yao J, Hu N, Ding F, Gu X. The protective effects of Achyranthes bidentata polypeptides in an experimental model of mouse sciatic nerve crush injury. Brain Res Bull. 2010;81:25–32
references.citation: Inserra MM, Bloch DA, Terris DJ. Functional indices for sciatic, peroneal, and posterior tibial nerve lesions in the mouse. Microsurgery. 1998;18:119–124
references.citation: Mohammadi R, Azizi S, Amini K. Effects of undifferentiated cultured omental adipose-derived stem cells on peripheral nerve regeneration. J Surg Res. 2013;180:e91–e97
references.citation: Batt JA, Bain JR. Tibial nerve transection: A standardized model for denervation-induced skeletal muscle atrophy in mice. J Vis Exp. 2013;3:e50657
references.citation: Amoh Y, Aki R, Hamada Y, et al. Nestin-positive hair follicle pluripotent stem cells can promote regeneration of impinged peripheral nerve injury. J Dermatol. 2012;39:33–38
references.citation: Lee EJ, Xu L, Kim GH, et al. Regeneration of peripheral nerves by transplanted sphere of human mesenchymal stem cells derived from embryonic stem cells. Biomaterials. 2012;33:7039–7046
references.citation: Liu F, Zhang C, Hoffman RM. Nestin-expressing stem cells from the hair follicle can differentiate into motor neurons and reduce muscle atrophy after transplantation to injured nerves. Tissue Eng Part A. 2014;20:656–662
references.citation: Nijhuis TH, de Boer SA, de Boer SA, et al. A new approach to assess the gastrocnemius muscle volume in rodents using ultrasound; comparison with the gastrocnemius muscle index. PLoS One. 2013;8:e54041
references.citation: Zhang J, Zhang G, Morrison B, Mori S, Sheikh KA. Magnetic resonance imaging of mouse skeletal muscle to measure denervation atrophy. Exp Neurol. 2008;212:448–457
references.citation: Jessen KR, Mirsky R. Origin and early development of Schwann cells. Microsc Res Tech. 1998;41:393–402
references.citation: Pereira Lopes FR, Frattini F, Marques SA, et al. Transplantation of bone-marrow-derived cells into a nerve guide resulted in transdifferentiation into Schwann cells and effective regeneration of transected mouse sciatic nerve. Micron. 2010;41:783–790
references.citation: Takeda K, Sowa Y, Nishino K, Itoh K, Fushiki S. Adipose-derived stem cells promote proliferation, migration, and tube formation of lymphatic endothelial cells in vitro by secreting lymphangiogenic factors. Ann Plast Surg. 2015;74:728–736
references.citation: Lopatina T, Kalinina N, Karagyaur M, et al. Adipose-derived stem cells stimulate regeneration of peripheral nerves: BDNF secreted by these cells promotes nerve healing and axon growth de novo. PLoS One. 2011;6:e17899
references.citation: Kingham PJ, Kolar MK, Novikova LN, Novikov LN, Wiberg M. Stimulating the neurotrophic and angiogenic properties of human adipose-derived stem cells enhances nerve repair. Stem Cells Dev. 2014;23:741–754
references.citation: Boerckel JD, Uhrig BA, Willett NJ, Huebsch N, Guldberg RE. Mechanical regulation of vascular growth and tissue regeneration in vivo. Proc Natl Acad Sci USA. 2011;108:E674–E680
references.citation: Hobson MI, Green CJ, Terenghi G. VEGF enhances intraneural angiogenesis and improves nerve regeneration after axotomy. J Anat. 2000;197:591–605
references.citation: Kosacka J, Nowicki M, Kacza J, Borlak J, Engele J, Spanel-Borowski K. Adipocyte-derived angiopoietin-1 supports neurite outgrowth and synaptogenesis of sensory neurons. J Neurosci Res. 2006;83:1160–1169
references.citation: Martini R, Fischer S, López-Vales R, David S. Interactions between Schwann cells and macrophages in injury and inherited demyelinating disease. Glia. 2008;56:1566–1577
references.citation: Santiago LY, Clavijo-Alvarez J, Brayfield C, Rubin JP, Marra KG. Delivery of adipose-derived precursor cells for peripheral nerve repair. Cell Transplant. 2009;18:145–158
references.citation: Suganuma S, Tada K, Hayashi K, et al. Uncultured adipose-derived regenerative cells promote peripheral nerve regeneration. J Orthop Sci. 2013;18:145–151
references.citation: Marconi S, Castiglione G, Turano E, et al. Human adipose-derived mesenchymal stem cells systemically injected promote peripheral nerve regeneration in the mouse model of sciatic crush. Tissue Eng Part A. 2012;18:1264–1272
references.citation: Tomita K, Madura T, Sakai Y, Yano K, Terenghi G, Hosokawa K. Glial differentiation of human adipose-derived stem cells: Implications for cell-based transplantation therapy. Neuroscience. 2013;236:55–65
references.citation: Brazelton TR, Blau HM. Optimizing techniques for tracking transplanted stem cells in vivo. Stem Cells. 2005;23:1251–1265
references.citation: Tabata Y. Biomaterial technology for tissue engineering applications. J R Soc Interface. 2009;6(Suppl 3):S311–S324