Mouse Degenerating Optic Axons Survived By Human Embryonic Stem Cell-Derived Neural Progenitor Cells

Document Type : Original Article


1 Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran

2 Ophthalmic Research Center, Tehran University of Medical Sciences, Tehran, Iran


Objective: Any damage to the optic nerve can potentially lead to degeneration of non-regenerating axons and ultimately
death of retinal ganglion cells (RGCs) that in most cases, are not curable by surgery or medication. Neuroprotective functions of different types of stem cells in the nervous system have been evaluated in many studies investigating the effectiveness of these cells in various retinal disease models. Neural progenitor cells (NPCs) secrete an assortment of trophic factors that are vital to the protection of the visual system. We aimed to assess the therapeutic potentials of NPCs in an ONC mouse model.
Materials and Methods: In this experimental study, NPCs were produced using noggin and retinoic acid from human embryonic stem cells (hESCs). Fifty mice were divided into the following three groups: i. Intact, ii. Vehicle [optic nerve crush+Hank’s balanced salt solution (HBSS)], and iii. Treatment (optic nerve crush+NPCs). The visual behavior of the mice was examined using the Visual Cliff test, and in terms of RGC numbers, they were assessed by Brn3a immunostaining and retrograde tracing using DiI injection.
Results: Intravenous injection of 50,000 NPCs through visual cliff did not produce any visual improvement. However, our data suggest that the RGCs protection was more than two-times in NPCs compared to the vehicle group as examined by Brn3a staining and retrograde tracing.
Conclusion: Our study indicated that intravenous injection of NPCs could protect RGCs probably mediated by trophic factors. Due to this ability and good manufacturing practices (GMP) grade production feasibility, NPCs may be used for optic nerve protection.


  1. Steinsapir KD, Seiff SR, Goldberg RA. Traumatic optic neuropathy: where do we stand? Ophthalmic Plast Reconstr Surg. 2002; 18(3): 232-234.
  2. Zuo KJ, Shi JB, Wen WP, Chen HX, Zhang XM, Xu G. Transnasal endoscopic optic nerve decompression for traumatic optic neuropathy: analysis of 155 cases. Zhonghua Yi Xue Za Zhi. 2009; 89(6): 389-392.
  3. Selhorst JB, Chen Y. The optic nerve. Semin Neurol. 2009; 29(1): 29-35.
  4. Zwart I, Hill AJ, Al-Allaf F, Shah M, Girdlestone J, Sanusi AB, et al. Umbilical cord blood mesenchymal stromal cells are neuroprotective and promote regeneration in a rat optic tract model. Exp Neurol. 2009; 216(2): 439-448.
  5. Aoki H, Hara A, Niwa M, Motohashi T, Suzuki T, Kunisada T. Transplantation of cells from eye-like structures differentiated from embryonic stem cells in vitro and in vivo regeneration of retinal ganglion- like cells. Graefes Arch Clin Exp Ophthalmol. 2008; 246(2): 255-265.
  6. Satarian L, Javan M, Kiani S, Hajikaram M, Mirnajafi-Zadeh J, Baharvand H. Engrafted human induced pluripotent stem cell-derived anterior specified neural progenitors protect the rat crushed optic nerve. PLoS One. 2013; 8(8): e71855.
  7. Mead B, Berry M, Logan A, Scott RA, Leadbeater W, Scheven BA. Stem cell treatment of degenerative eye disease. Stem Cell Res. 2015; 14(3): 243-257.
  8. Mollamohammadi S, Taei A, Pakzad M, Totonchi M, Seifinejad A, Masoudi N, et al. A simple and efficient cryopreservation method for feeder-free dissociated human induced pluripotent stem cells and human embryonic stem cells. Hum Reprod. 2009; 24(10): 2468- 2476.
  9. Seyedrazizadeh SZ, Poosti S, Nazari A, Alikhani M, Shekari F, Pakdel F, et al. Extracellular vesicles derived from human ES-MSCs protect retinal ganglion cells and preserve retinal function in a rodent model of optic nerve injury. Stem Cell Res Ther. 2020; 11(1): 203.
  10. de Lima S, Koriyama Y, Kurimoto T, Oliveira JT, Yin Y, Li Y, et al. Full-length axon regeneration in the adult mouse optic nerve and partial recovery of simple visual behaviors. Proc Natl Acad Sci USA. 2012; 109(23): 9149-9154.
  11. Kokaia Z, Martino G, Schwartz M, Lindvall O. Cross-talk between neural stem cells and immune cells: the key to better brain repair? Nat Neurosci. 2012; 15(8): 1078-1087.
  12. Drago D, Cossetti C, Iraci N, Gaude E, Musco G, Bachi A, et al. The stem cell secretome and its role in brain repair. Biochimie. 2013; 95(12): 2271-2285.
  13. Blurton-Jones M, Kitazawa M, Martinez-Coria H, Castello NA, Müller FJ, Loring JF, et al. Neural stem cells improve cognition via BDNF in a transgenic model of alzheimer disease. Proc Natl Acad Sci USA. 2009; 106(32): 13594-13599.
  14. Kerkis I, Haddad MS, Valverde CW, Glosman S. Neural and mesenchymal stem cells in animal models of huntington’s disease: past experiences and future challenges. Stem Cell Res Ther. 2015; 6: 232.
  15. Abe K. Therapeutic potential of neurotrophic factors and neural stem cells against ischemic brain injury. J Cereb Blood Flow Metab. 2000; 20(10): 1393-1408.
  16. De Feo D, Merlini A, Laterza C, Martino G. Neural stem cell transplantation in central nervous system disorders: from cell replacement to neuroprotection. Curr Opin Neurol. 2012; 25(3): 322-33.
  17. Leaver SG, Cui Q, Plant GW, Arulpragasam A, Hisheh S, Verhaagen J, et al. AAV-mediated expression of CNTF promotes longterm survival and regeneration of adult rat retinal ganglion cells. Gene Ther. 2006; 13(18): 1328-1341.
  18. Johnson TV, DeKorver NW, Levasseur VA, Osborne A, Tassoni A, Lorber B, et al. Identification of retinal ganglion cell neuroprotection conferred by platelet-derived growth factor through analysis of the mesenchymal stem cell secretome. Brain. 2014; 137(Pt 2): 503-519.
  19. Sahu A, Foulsham W, Amouzegar A, Mittal SK, Chauhan SK. The therapeutic application of mesenchymal stem cells at the ocular surface. Ocul Surf. 2019; 17(2): 198-207.
  20. Tang Y, Yu P, Cheng L. Current progress in the derivation and therapeutic application of neural stem cells. Cell Death Dis. 2017; 8(10): e3108.
  21. Banin E, Obolensky A, Idelson M, Hemo I, Reinhardtz E, Pikarsky E, et al. Retinal incorporation and differentiation of neural precursors derived from human embryonic stem cells. Stem Cells. 2006; 24(2): 246-257.
  22. Xu L, Yan J, Chen D, Welsh AM, Hazel T, Johe K, et al. Human neural stem cell grafts ameliorate motor neuron disease in SOD-1 transgenic rats. Transplantation. 2006; 82(7): 865-875.
  23. Tan J, Zheng X, Zhang S, Yang Y, Wang X, Yu X, et al. Response of the sensorimotor cortex of cerebral palsy rats receiving transplantation of vascular endothelial growth factor 165-transfected neural stem cells. Neural Regen Res. 2014; 9(19): 1763-1769.
  24. Salehi H, Karbalaie K, Salamian A, Kiani A, Razavi S, Nasr-Esfahani MH, et al. Differentiation of human ES cell-derived neural progenitors to neuronal cells with regional specific identity by coculturing of notochord and somite. Stem Cell Res. 2012; 8(1): 120- 133.
  25. Shaham O, Menuchin Y, Farhy C, Ashery-Padan R. Pax6: a multilevel regulator of ocular development. Prog Retin Eye Res. 2012; 31(5): 351-376.
  26. Mu X, Klein WH. A gene regulatory hierarchy for retinal ganglion cell specification and differentiation. Semin Cell Dev Biol. 2004; 15(1): 115-123.
  27. Gamm DM, Wang S, Lu B, Girman S, Holmes T, Bischoff N, et al. Protection of visual functions by human neural progenitors in a rat model of retinal disease. PLoS One. 2007; 2(3): e338.
  28. Liang YX, Yang J, Yuan TF, So KF. Uptake of retrograde tracers by intact optic nerve axons: a new way to label retinal ganglion cells. PLoS One. 2015; 10(6): e0128718.