Cell J, Vol 24, No 3, March 2022 120 Original Article Mouse Degenerating Optic Axons Survived by Human Embryonic Stem Cell-Derived Neural Progenitor Cells Shiva Nemati, Ph.D. 1 , Zahra Seiedrazizadeh, M.Sc. 1 , Susan Simorgh, M.Sc. 1 , Mahdi Hesaraki, M.Sc. 1 , Sahar Kiani, Ph.D. 1 , Mohammad Javan, Ph.D. 1, 2 , Farzad Pakdel, M.D. 3 , Leila Satarian, Ph.D. 1 * 1. Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran 2. Department of Physiology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran 3. Ophthalmic Research Center, Tehran University of Medical Sciences, Tehran, Iran *Corresponding Address: P.O.Box: 16635-148, Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran Email: l.satarian@royan-rc.ac.ir Received: 19/November/2020, Accepted: 24/January/2021 Abstract 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. Keywords: Human Embryonic Stem Cells, Optic Nerve Injury, Visual Cliff Cell Journal (Yakhteh), Vol 24, No 3, March 2022, Pages: 120- 126 Citation: Nemati Sh, Seiedrazizadeh Z, Simorgh S, Hesaraki M, Kiani S, Javan M, Pakdel F, Satarian L. Mouse degenerating optic axons survived by human embryonic stem cell-derived neural progenitor cells . Cell J. 2022; 24(3): 120-126. doi: 10.22074/cellj.2022.7873. This open-access article has been published under the terms of the Creative Commons Attribution Non-Commercial 3.0 (CC BY-NC 3.0). Introduction Nearly 0.5-5 percent of vehicular accidents lead to optic nerve crush (ONC) injuries; serious damages that could lead to cell degradation and eventual vision loss, due to the limitations in retinal ganglion cells (RGCs) regeneration (1). Currently available medical interventions involving administration of neuroprotective medications such as corticosteroids to reduce infammation, or surgery to remove pressure, have yielded little therapeutic success (2). Therefore, a large number of injured individuals- mostly of young ages-suffer from blindness (3). Nevertheless, it is anticipated that stem cells, which have the potential to cure neurological disorders, may help in overcoming this issue (4). The following therapeutic methods are currently employed for neuropathological conditions: protecting the damaged cells, preventing further degeneration, and replacing the degenerated cells with cell transplants. RGC axons transfer the signals induced by visual stimuli in the eye to the brain’s targets. Since RGC axons are very long and possess complex pathways, it does not seem logical to replace the degenerated cells with cell transplants. However, protecting the degenerating RGCs might be a promising approach. Due to the protective and regenerative properties of stem cells, various types of these cells, including adult, embryonic and induced pluripotent stem cells at different levels of differentiation, have been studied in a variety of retinal disease models (5). NPCs are located in the adult brain or derivatives from pluripotent stem cells. In the adult brain, they are found in two defned areas named subventricular zone (SVZ), which is around the ventricles of cerebral cortex, and subgranular zone (SGZ), located in the hippocampus. These parts of the brain are in charge of generating new neural cells. An injury or disease leading to neuronal loss and infammation in the adult CNS will activate the NPCs by increasing their proliferation and migration rates. Studies have demonstrated that NPCs act mostly through