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Enrichment of retinal ganglion and Müller glia progenitors from retinal organoids derived from human induced pluripotent stem cells-possibilities and current limitations

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World Journal of Stem Cells 2020年第10期12卷 1171-1183页

作者： Kristine Karla Freude Sarkis Saruhanian Alanna McCauley Colton Paterson Madeleine Odette Annika Oostenink Poul Hyttel Mark Gillies Henriette Haukedal Miriam Kolko Department of Veterinary and Animal Sciences Group of Stem Cell Models for Studies of Neurodegenerative DiseasesSection for Pathobiological SciencesFaculty of Health and Medical SciencesUniversity of CopenhagenFrederiksberg 1870Denmark Department of Drug Design and Pharmacology University of CopenhagenCopenhagen 2100Denmark Save Sight Institute South BlockSydney Eye HospitalSydney 2000Australia Department of Ophthalmology Copenhagen University HospitalRigshospitalet-GlostrupGlostrup 2600Denmark

BACKGROUND retinal organoids serve as excellent human-specific disease models for conditions affecting otherwise inaccessible retinal tissue from *** permit the isolation of key cell types affected in various eye diseases including retinal ganglion cells(RGCs)and Müller *** To refine human-induced pluripotent stem cells(hiPSCs)differentiated into threedimensional(3D)retinal organoids to generate sufficient numbers of RGCs and Müller glia progenitors for downstream *** In this study we described,evaluated,and refined methods with which to generate Müller glia and RGC progenitors,isolated them via magnetic-activated cell sorting,and assessed their lineage stability after prolonged 2D *** progenitor populations were characterized via quantitative PCR and immunocytochemistry,and the ultrastructural composition of retinal organoid cells was *** Our study confirms the feasibility of generating marker-characterized Müller glia and RGC progenitors within retinal *** retinal organoids can be dissociated and the Müller glia and RGC progenitor-like cells isolated via magnetic-activated cell sorting and propagated as *** Enrichment of Müller glia and RGC progenitors from retinal organoids is a feasible method with which to study cell type-specific disease phenotypes and to potentially generate specific retinal populations for cell replacement therapies.

关键词： Human induced pluripotent stem cells retinal organoids retinal ganglion cells Müller glia Progenitors Cell-type enrichment

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Cell replacement with stem cell-derived retinal ganglion cells from different protocols

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Neural Regeneration Research 2024年第4期19卷 807-810页

作者： Ziming Luo Kun-Che Chang Spencer Center for Vision Research Byers Eye InstituteStanford University School of Medicine Department of Ophthalmology Louis J.Fox Center for Vision RestorationUniversity of Pittsburgh School of Medicine Department of Neurobiology Center of NeuroscienceUniversity of Pittsburgh School of Medicine

Glaucoma,characterized by a degenerative loss of retinal ganglion cells,is the second leading cause of blindness *** is currently no cure for vision loss in glaucoma because retinal ganglion cells do not regenerate and are not replaced after *** stem cell-derived retinal ganglion cell transplant is a potential therapeutic strategy for retinal ganglion cell degenerative *** this review,we first discuss a 2D protocol for retinal ganglion cell differentiation from human stem cell culture,including a rapid protocol that can generate retinal ganglion cells in less than two weeks and focus on their transplantation ***,we discuss using 3D retinal organoids for retinal ganglion cell transplantation,comparing cell suspensions and *** review provides insight into current knowledge on human stem cell-derived retinal ganglion cell differentiation and transplantation,with an impact on the field of regenerative medicine and especially retinal ganglion cell degenerative diseases such as glaucoma and other optic neuropathies.

关键词： cell clumps cell suspension cell transplantation differentiation direct-induced protocol glaucoma optic neuropathy regenerative medicine retinal ganglion cell retinal organoids stem cells

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International Journal of Ophthalmology(English edition) 2021年第8期14卷 1138-1150页

作者： Hao-Jue Xu Qi-You Li Ting Zou Zheng-Qin Yin Southwest Hospital/Southwest Eye Hospital Third Military Medical University(Army Medical University)Chongqing 400038China Key Lab of Visual Damage and Regeneration&Restoration of Chongqing Chongqing 400038China

AIM:To explore the temporal mitochondrial characteristics of retinal pigment epithelium(RPE)cells obtained from human embryonic stem cells(hESC)-derived retinal organoids(hEROs-RPE),to verify the optimal period for using hEROs-RPE as donor cells from the aspect of mitochondria and to optimize RPE cell-based therapeutic strategies for age-related macular degeneration(AMD).METHODS:RPE cells were obtained from hEROs and from spontaneous differentiation(SD-RPE).The mitochondrial characteristics were analyzed every 20 d from day 60 to *** quantity was measured by MitoTracker Green *** electron microscopy(TEM)was adopted to assess the morphological features of the mitochondria,including their distribution,length,and *** membrane potentials(MMPs)were determined by JC-1 staining and evaluated by flow cytometry,reactive oxygen species(ROS)levels were evaluated by flow cytometry,and adenosine triphosphate(ATP)levels were measured by a *** between two groups were analyzed by the independentsamples t-test,and comparisons among multiple groups were made using one-way ANOVA or Kruskal-Wallis H test when equal variance was not ***:hEROs-RPE and SD-RPE cells from day 60 to 160 were successfully differentiated from hESCs and expressed RPE markers(Pax6,MITF,Bestrophin-1,RPE65,Cralbp).RPE features,including a cobblestonelike morphology with tight junctions(ZO-1),pigments and microvilli,were also observed in both hEROs-RPE and SDRPE *** mitochondrial quantities of hEROs-RPE and SD-RPE cells both peaked at day ***,the cristae of hEROs-RPE mitochondria were less mature and abundant than those of SD-RPE mitochondria at day 80,with hEROsRPE mitochondria becoming mature at day *** hEROs-RPE and SD-RPE cells showed low ROS levels from day 100 to 140 and maintained a normal MMP during this ***,hEROs-RPE mitochondria maintained a longer time to produce high levels of ATP(from day 120

关键词： mitochondria retinal organoids retinal pigment epithelium cells human embryonic stem cells,retinal degenerative diseases

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