Schwann cell-free adult canine olfactory ensheathing cell preparations from olfactory bulb and mucosa display differential migratory and neurite growth-promoting properties in vitro

BMC Neuroscience

Gerd Bicker²  Konstantin Wewetzer⁴  Wolfgang Baumgärtner²  Susanne Ziege³  Frank Roloff¹ 

[1] Division of Cell Biology, University of Veterinary Medicine Hannover, Bischofsholer Damm 15/102, 30173 Hannover, Germany;Center for Systems Neuroscience Hannover, Hannover, Germany;Department of Pathology, University of Veterinary Medicine Hannover, Bünteweg 17, 30559 Hannover, Germany;Department of Functional and Applied Anatomy, Hannover Medical School, Carl-Neuberg-Str.1, 30625 Hannover, Germany

关键词: Regeneration;    Human NT-2 neurons;    Large animal model;    Scratch wound assay;    Glia;   

Others  :  1131322
DOI  :  10.1186/1471-2202-14-141

 received in 2013-05-23, accepted in 2013-11-07,  发布年份 2013

【 摘 要 】

Background

Transplantation of olfactory ensheathing cells (OEC) and Schwann cells (SC) is a promising therapeutic strategy to promote axonal growth and remyelination after spinal cord injury. Previous studies mainly focused on the rat model though results from primate and porcine models differed from those in the rat model. Interestingly, canine OECs show primate-like in vitro characteristics, such as absence of early senescence and abundance of stable p75^NTR expression indicating that this species represents a valuable translational species for further studies. So far, few investigations have tested different glial cell types within the same study under identical conditions. This makes it very difficult to evaluate contradictory or confirmatory findings reported in various studies. Moreover, potential contamination of OEC preparations with Schwann cells was difficult to exclude. Thus, it remains rather controversial whether the different glial types display distinct cellular properties.

Results

Here, we established cultures of Schwann cell-free OECs from olfactory bulb (OB-OECs) and mucosa (OM-OECs) and compared them in assays to Schwann cells. These glial cultures were obtained from a canine large animal model and used for monitoring migration, phagocytosis and the effects on in vitro neurite growth. OB-OECs and Schwann cells migrated faster than OM-OECs in a scratch wound assay. Glial cell migration was not modulated by cGMP and cAMP signaling, but activating protein kinase C enhanced motility. All three glial cell types displayed phagocytic activity in a microbead assay. In co-cultures with of human model (NT2) neurons neurite growth was maximal on OB-OECs.

Conclusions

These data provide evidence that OB- and OM-OECs display distinct migratory behavior and interaction with neurites. OB-OECs migrate faster and enhance neurite growth of human model neurons better than Schwann cells, suggesting distinct and inherent properties of these closely-related cell types. Future studies will have to address whether, and how, these cellular properties correlate with the in vivo behavior after transplantation.

【 授权许可】

   
2013 Roloff et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Bradbury EJ, McMahon SB: Spinal cord repair strategies: why do they work? Nat Rev Neurosci 2006, 7:644-653.
• [2]Kim J, Schafer J, Ming G: New directions in neuroregeneration. Expert Opin Biol Ther 2006, 6:735-738.
• [3]Harrop JS, Hashimoto R, Norvell D, Raich A, Aarabi B, Grossman RG, Guest JD, Tator CH, Chapman J, Fehlings MG: Evaluation of clinical experience using cell-based therapies in patients with spinal cord injury: a systematic review. J Neurosurg Spine 2012, 17(1 Suppl):230-246.
• [4]Tabakow P, Jarmundowicz W, Czapiga B, Fortuna W, Miedzybrodzki R, Czyz M, Huber J, Szarek D, Okurowski S, Szewczyk P, Gorski A, Raisman G: Transplantation of autologous olfactory ensheathing cells in complete human spinal cord injury. Cell Transplant 2013, 22(9):1591-1612.
• [5]Granger N, Blamires H, Franklin RJM, Jeffery ND: Autologous olfactory mucosal cell transplants in clinical spinal cord injury: a randomized double-blinded trial in a canine translational model. Brain 2012, 135(Pt 11):3227-3237.
• [6]Li Y: Repair of Adult Rat Corticospinal Tract by Transplants of Olfactory Ensheathing Cells. Science 1997, 277:2000-2002.
• [7]Ramón-Cueto A, Cordero MI, Santos-Benito FF, Avila J: Functional recovery of paraplegic rats and motor axon regeneration in their spinal cords by Olfactory Ensheathing Glia. Neuron 2000, 25:425-435.
• [8]Li Y, Decherchi P, Raisman G: Transplantation of olfactory ensheathing cells into spinal cord lesions restores breathing and climbing. J Neurosci 2003, 23:727-731.
• [9]Ziegler MD, Hsu D, Takeoka A, Zhong H, Ramón-Cueto A, Phelps PE, Roy RR, Edgerton VR: Further evidence of olfactory ensheathing glia facilitating axonal regeneration after a complete spinal cord transection. Exp Neurol 2011, 229:109-119.
• [10]Ramón-Cueto A, Plant GW, Avila J, Bunge MB: Long-distance axonal regeneration in the transected adult rat spinal cord is promoted by olfactory ensheathing glia transplants. J Neurosci 1998, 18:3803-3815.
• [11]Baron-Van Evercooren A, Avellana-Adalid V, Ben Younes-Chennoufi A, Gansmuller A, Nait-Oumesmar B, Vignais L: Cell-cell interactions during the migration of myelin-forming cells transplanted in the demyelinated spinal cord. Glia 1996, 16:147-164.
• [12]Li Y, Raisman G: Integration of transplanted cultured Schwann cells into the long myelinated fiber tracts of the adult spinal cord. Exp Neurol 1997, 145(2 Pt 1):397-411.
• [13]Dibaj P, Nadrigny F, Steffens H, Scheller A, Hirrlinger J, Schomburg ED, Neusch C, Kirchhoff F: NO mediates microglial response to acute spinal cord injury under ATP control in vivo. Glia 2010, 58:1133-1144.
• [14]Huo Y, Yin X-L, Ji S-X, Zou H, Lang M, Zheng Z, Cai X-F, Liu W, Chen C-L, Zhou Y-G, Yuan R-D, Ye J: Inhibition of retinal ganglion cell axonal outgrowth through the amino-nogo-a signaling pathway. Neurochem Res 2013, 38(7):1365-1374.
• [15]Wewetzer K, Kern N, Ebel C, Radtke C, Brandes G: Phagocytosis of O4+ axonal fragments in vitro by p75- neonatal rat olfactory ensheathing cells. Glia 2005, 49:577-587.
• [16]Panni P, Ferguson IA, Beacham I, Mackay-Sim A, Ekberg JAK, St John JA: Phagocytosis of bacteria by olfactory ensheathing cells and Schwann cells. Neurosci Lett 2013, 539:65-70.
• [17]Su Z, Chen J, Qiu Y, Yuan Y, Zhu F, Zhu Y, Liu X, Pu Y, He C: Olfactory ensheathing cells: The primary innate immunocytes in the olfactory pathway to engulf apoptotic olfactory nerve debris. Glia 2013, 61:490-503.
• [18]Ziege S, Baumgärtner W, Wewetzer K: Toward defining the regenerative potential of olfactory mucosa: establishment of schwann cell-free adult canine olfactory ensheathing cell preparations suitable for transplantation. Cell Transplant 2013, 22:355-367.
• [19]Horner PJ, Gage FH: Regenerating the damaged central nervous system. Nature 2000, 407:963-970.
• [20]Song H, Poo M: The cell biology of neuronal navigation. Nat Cell Biol 2001, 3:E81-E88.
• [21]Schwartz RE, Linehan JL, Painschab MS, Hu W-S, Verfaillie CM, Kaufman DS: Defined conditions for development of functional hepatic cells from human embryonic stem cells. Stem Cells Dev 2005, 14:643-655.
• [22]Przyborski SA, Morton IE, Wood A, Andrews PW: Developmental regulation of neurogenesis in the pluripotent human embryonal carcinoma cell line NTERA-2. Eur J Neurosci 2000, 12:3521-3528.
• [23]Houldsworth J, Heath SC, Bosl GJ, Studer L, Chaganti RSK: Expression profiling of lineage differentiation in pluripotential human embryonal carcinoma cells. Cell Growth Differ 2002, 13:257-264.
• [24]Przyborski SA, Smith S, Wood A: Transcriptional profiling of neuronal differentiation by human embryonal carcinoma stem cells in vitro. Stem Cells 2003, 21:459-471.
• [25]Smith B, Treadwell J, Zhang D, Ly D, McKinnell I, Walker PR, Sikorska M: Large-scale expression analysis reveals distinct microRNA profiles at different stages of human neurodevelopment. PLoS ONE 2010, 5:e11109.
• [26]Meintanis S, Thomaidou D, Jessen KR, Mirsky R, Matsas R: The neuron-glia signal beta-neuregulin promotes Schwann cell motility via the MAPK pathway. Glia 2001, 34:39-51.
• [27]Liang C-C, Park AY, Guan J-L: In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc 2007, 2:329-333.
• [28]Tegenge MA, Bicker G: Nitric oxide and cGMP signal transduction positively regulates the motility of human neuronal precursor (NT2) cells. J Neurochem 2009, 110:1828-1841.
• [29]Weeraratna AT, Jiang Y, Hostetter G, Rosenblatt K, Duray P, Bittner M, Trent JM: Wnt5a signaling directly affects cell motility and invasion of metastatic melanoma. Cancer Cell 2002, 1:279-288.
• [30]Moors M, Cline JE, Abel J, Fritsche E: ERK-dependent and -independent pathways trigger human neural progenitor cell migration. Toxicol Appl Pharmacol 2007, 221:57-67.
• [31]Yamamoto H, Kitadai Y, Yamamoto H, Oue N, Ohdan H, Yasui W, Kikuchi A: Laminin gamma2 mediates Wnt5a-induced invasion of gastric cancer cells. Gastroenterology 2009, 137:242-252. 252.e1–6
• [32]Sumagin R, Robin AZ, Nusrat A, Parkos CA: Activation of PKCβII by PMA Facilitates Enhanced Epithelial Wound Repair through Increased Cell Spreading and Migration. PLoS ONE 2013, 8:e55775.
• [33]Windus LCE, Chehrehasa F, Lineburg KE, Claxton C, Mackay-Sim A, Key B, St John JA: Stimulation of olfactory ensheathing cell motility enhances olfactory axon growth. Cell Mol Life Sci 2011, 68:3233-3247.
• [34]Leung JYK, Chapman JA, Harris JA, Hale D, Chung RS, West AK, Chuah MI: Olfactory ensheathing cells are attracted to, and can endocytose, bacteria. Cell Mol Life Sci 2008, 65:2732-2739.
• [35]Tegenge MA, Roloff F, Bicker G: Rapid differentiation of human embryonal carcinoma stem cells (NT2) into neurons for neurite outgrowth analysis. Cell Mol Neurobiol 2011, 31:635-643.
• [36]Doucette R: Olfactory ensheathing cells: potential for glial cell transplantation into areas of CNS injury. Histol Histopathol 1995, 10:503-507.
• [37]Barnett SC, Chang L: Olfactory ensheathing cells and CNS repair: going solo or in need of a friend? Trends Neurosci 2004, 27:54-60.
• [38]Richter MW, Roskams AJ: Olfactory ensheathing cell transplantation following spinal cord injury: hype or hope? Exp Neurol 2008, 209:353-367.
• [39]Guntinas-Lichius O, Wewetzer K, Tomov TL, Azzolin N, Kazemi S, Streppel M, Neiss WF, Angelov DN: Transplantation of olfactory mucosa minimizes axonal branching and promotes the recovery of vibrissae motor performance after facial nerve repair in rats. J Neurosci 2002, 22:7121-7131.
• [40]Li Y, Yamamoto M, Raisman G, Choi D, Carlstedt T: An experimental model of ventral root repair showing the beneficial effect of transplanting olfactory ensheathing cells. Neurosurgery 2007, 60:734-740. discussion 740–741
• [41]Radtke C, Vogt PM: Peripheral nerve regeneration: a current perspective. Eplasty 2009, 9:e47.
• [42]López-Vales R, García-Alías G, Forés J, Navarro X, Verdú E: Increased expression of cyclo-oxygenase 2 and vascular endothelial growth factor in lesioned spinal cord by transplanted olfactory ensheathing cells. J Neurotrauma 2004, 21:1031-1043.
• [43]Richter MW, Fletcher PA, Liu J, Tetzlaff W, Roskams AJ: Lamina propria and olfactory bulb ensheathing cells exhibit differential integration and migration and promote differential axon sprouting in the lesioned spinal cord. J Neurosci 2005, 25:10700-10711.
• [44]Pearse DD, Sanchez AR, Pereira FC, Andrade CM, Puzis R, Pressman Y, Golden K, Kitay BM, Blits B, Wood PM, Bunge MB: Transplantation of Schwann cells and/or olfactory ensheathing glia into the contused spinal cord: Survival, migration, axon association, and functional recovery. Glia 2007, 55:976-1000.
• [45]Chehrehasa F, Windus LCE, Ekberg JAK, Scott SE, Amaya D, Mackay-Sim A, St John JA: Olfactory glia enhance neonatal axon regeneration. Mol Cell Neurosci 2010, 45:277-288.
• [46]Liu Q, Yu H-M, Dai C-F, Li W, Zhu Y-Y, Gu Y-R, Li H-W: Olfactory ensheathing cells promote the survival of newborn rat spiral ganglion cells in vitro. Sheng Li Xue Bao 2010, 62:115-121.
• [47]Coutts DJ, Humphries CE, Zhao C, Plant GW, Franklin RJ: Embryonic-derived olfactory ensheathing cells remyelinate focal areas of spinal cord demyelination more efficiently than neonatal or adult-derived cells. Cell Transplant 2013, 22(7):1249-61.
• [48]Techangamsuwan S, Imbschweiler I, Kreutzer R, Kreutzer M, Baumgärtner W, Wewetzer K: Similar behaviour and primate-like properties of adult canine Schwann cells and olfactory ensheathing cells in long-term culture. Brain Res 2008, 1240:31-38.
• [49]Radtke C, Lankford KL, Wewetzer K, Imaizumi T, Fodor WL, Kocsis JD: Impaired spinal cord remyelination by long-term cultured adult porcine olfactory ensheathing cells correlates with altered in vitro phenotypic properties. Xenotransplantation 2010, 17:71-80.
• [50]Wewetzer K, Radtke C, Kocsis J, Baumgärtner W: Species-specific control of cellular proliferation and the impact of large animal models for the use of olfactory ensheathing cells and Schwann cells in spinal cord repair. Exp Neurol 2011, 229:80-87.
• [51]Huang H, Xi H, Chen L, Zhang F, Liu Y: Long-term outcome of olfactory ensheathing cell therapy for patients with complete chronic spinal cord injury. Cell Transplant 2012, 21(Suppl 1):S23-S31.
• [52]Lankford KL, Sasaki M, Radtke C, Kocsis JD: Olfactory ensheathing cells exhibit unique migratory, phagocytic, and myelinating properties in the X-irradiated spinal cord not shared by Schwann cells. Glia 2008, 56:1664-1678.
• [53]Lu P, Yang H, Culbertson M, Graham L, Roskams AJ, Tuszynski MH: Olfactory ensheathing cells do not exhibit unique migratory or axonal growth-promoting properties after spinal cord injury. J Neurosci 2006, 26:11120-11130.
• [54]Windus LCE, Claxton C, Allen CL, Key B, St John JA: Motile membrane protrusions regulate cell-cell adhesion and migration of olfactory ensheathing glia. Glia 2007, 55:1708-1719.
• [55]Ramer LM, Richter MW, Roskams AJ, Tetzlaff W, Ramer MS: Peripherally-derived olfactory ensheathing cells do not promote primary afferent regeneration following dorsal root injury. Glia 2004, 47:189-206.
• [56]Su Z, Yuan Y, Chen J, Cao L, Zhu Y, Gao L, Qiu Y, He C: Reactive astrocytes in glial scar attract olfactory ensheathing cells migration by secreted TNF-alpha in spinal cord lesion of rat. PLoS ONE 2009, 4:e8141.
• [57]Shen Y, Qian Y, Zhang H, Zuo B, Lu Z, Fan Z, Zhang P, Zhang F, Zhou C: Guidance of olfactory ensheathing cell growth and migration on electrospun silk fibroin scaffolds. Cell Transplant 2010, 19:147-157.
• [58]Simón D, Martín-Bermejo MJ, Gallego-Hernández MT, Pastrana E, García-Escudero V, García-Gómez A, Lim F, Díaz-Nido J, Avila J, Moreno-Flores MT: Expression of plasminogen activator inhibitor-1 by olfactory ensheathing glia promotes axonal regeneration. Glia 2011, 59:1458-1471.
• [59]Cai F, Campana WM, Tomlinson DR, Fernyhough P: Transforming growth factor-beta1 and glial growth factor 2 reduce neurotrophin-3 mRNA expression in cultured Schwann cells via a cAMP-dependent pathway. Brain Res Mol Brain Res 1999, 71:256-264.
• [60]Vincent AJ, West AK, Chuah MI: Morphological plasticity of olfactory ensheathing cells is regulated by cAMP and endothelin-1. Glia 2003, 41:393-403.
• [61]Wang Y, Huang Z-H: cAMP mediates the morphological change of cultured olfactory ensheathing cells induced by serum. Sheng Li Xue Bao 2011, 63:31-38.
• [62]Bove PF, Wesley UV, Greul A-K, Hristova M, Dostmann WR, van der Vliet A: Nitric oxide promotes airway epithelial wound repair through enhanced activation of MMP-9. Am J Respir Cell Mol Biol 2007, 36:138-146.
• [63]Tegenge MA, Rockel TD, Fritsche E, Bicker G: Nitric oxide stimulates human neural progenitor cell migration via cGMP-mediated signal transduction. Cell Mol Life Sci 2011, 68:2089-2099.
• [64]Harris JA, West AK, Chuah MI: Olfactory ensheathing cells: nitric oxide production and innate immunity. Glia 2009, 57:1848-1857.
• [65]Buss A, Pech K, Merkler D, Kakulas BA, Martin D, Schoenen J, Noth J, Schwab ME, Brook GA: Sequential loss of myelin proteins during Wallerian degeneration in the human spinal cord. Brain 2005, 128(Pt 2):356-364.
• [66]David S, Fry EJ, López-Vales R: Novel roles for Nogo receptor in inflammation and disease. Trends Neurosci 2008, 31:221-226.
• [67]Huo SJ, Li YC, Xie J, Li Y, Raisman G, Zeng YX, He JR, Weng CH, Yin ZQ: Transplanted olfactory ensheathing cells reduce retinal degeneration in royal college of surgeons rats. Curr Eye Res 2012, 37:749-758.
• [68]Li Y, Field PM, Raisman G: Olfactory ensheathing cells and olfactory nerve fibroblasts maintain continuous open channels for regrowth of olfactory nerve fibres. Glia 2005, 52:245-251.
• [69]Omar M, Bock P, Kreutzer R, Ziege S, Imbschweiler I, Hansmann F, Peck C-T, Baumgärtner W, Wewetzer K: Defining the morphological phenotype: 2′,3′-cyclic nucleotide 3′-phosphodiesterase (CNPase) is a novel marker for in situ detection of canine but not rat olfactory ensheathing cells. Cell Tissue Res 2011, 344:391-405.
• [70]Ziege S, Wewetzer K, Schöne K, Baumgärtner W: Special neurite growth-promoting capacity and growth factor responsiveness of nasal mucosa-derived olfactory ensheathing cells unraveled by Schwann cell depletion. Exp Neurol 2013.
• [71]Bradbury EJ, McMahon SB, Ramer MS: Keeping in touch: sensory neurone regeneration in the CNS. Trends Pharmacol Sci 2000, 21:389-394.
• [72]Neumann S, Woolf CJ: Regeneration of dorsal column fibers into and beyond the lesion site following adult spinal cord injury. Neuron 1999, 23:83-91.
• [73]Fudge NJ, Mearow KM: Extracellular matrix-associated gene expression in adult sensory neuron populations cultured on a laminin substrate. BMC Neurosci 2013, 14:15. BioMed Central Full Text
• [74]Ramón-Cueto A, Nieto-Sampedro M: Regeneration into the spinal cord of transected dorsal root axons is promoted by ensheathing glia transplants. Exp Neurol 1994, 127:232-244.
• [75]Paquet-Durand F, Bicker G: Human model neurons in studies of brain cell damage and neural repair. Curr Mol Med 2007, 7:541-554.
• [76]Podrygajlo G, Tegenge MA, Gierse A, Paquet-Durand F, Tan S, Bicker G, Stern M: Cellular phenotypes of human model neurons (NT2) after differentiation in aggregate culture. Cell Tissue Res 2009, 336:439-452.
• [77]Podrygajlo G, Song Y, Schlesinger F, Krampfl K, Bicker G: Synaptic currents and transmitter responses in human NT2 neurons differentiated in aggregate culture. Neurosci Lett 2010, 468:207-210.
• [78]Blackmore M, Letourneau PC: Changes within maturing neurons limit axonal regeneration in the developing spinal cord. J Neurobiol 2006, 66:348-360.
• [79]Qiu J, Cai D, Dai H, McAtee M, Hoffman PN, Bregman BS, Filbin MT: Spinal axon regeneration induced by elevation of cyclic AMP. Neuron 2002, 34:895-903.
• [80]Gensel JC, Kigerl KA, Mandrekar-Colucci SS, Gaudet AD, Popovich PG: Achieving CNS axon regeneration by manipulating convergent neuro-immune signaling. Cell Tissue Res 2012, 349:201-213.
• [81]Ramer LM, Au E, Richter MW, Liu J, Tetzlaff W, Roskams AJ: Peripheral olfactory ensheathing cells reduce scar and cavity formation and promote regeneration after spinal cord injury. J Comp Neurol 2004, 473:1-15.
• [82]Toft A, Tome M, Barnett SC, Riddell JS: A comparative study of glial and non-neural cell properties for transplant-mediated repair of the injured spinal cord. Glia 2013, 61:513-528.
• [83]Wewetzer K, Grothe C, Christ B, Seilheimer B: Identification and characterization of differentiation-dependent Schwann cell surface antigens by novel monoclonal antibodies: introduction of a marker common to the non-myelin-forming phenotype. Glia 1997, 19:213-226.