scholarly journals Human Retinal Progenitor Cells Derived Small Extracellular Vesicles Delay Retinal Degeneration: A Paradigm for Cell-free Therapy

2021 ◽  
Vol 12 ◽  
Author(s):  
Min Chen ◽  
Chunge Ren ◽  
Bangqi Ren ◽  
Yajie Fang ◽  
Qiyou Li ◽  
...  
Keyword(s):  
Retinal Degeneration ◽  
Progenitor Cells ◽  
Extracellular Vesicles ◽  
Target Genes ◽  
Outer Nuclear Layer ◽  
Subretinal Space ◽  
Retinal Progenitor Cells ◽  
Retinal Progenitor ◽  
Rcs Rats

Retinal degeneration is a leading cause of irreversible vision impairment and blindness worldwide. Previous studies indicate that subretinal injection of human retinal progenitor cells (hRPCs) can delay the progression of retinal degeneration, preserve retinal function, and protect photoreceptor cells from death, albeit the mechanism is not well understood. In this study, small extracellular vesicles derived from hRPCs (hRPC-sEVs) were injected into the subretinal space of retinal dystrophic RCS rats. We find that hRPC-sEVs significantly preserve the function of retina and thickness of the outer nuclear layer (ONL), reduce the apoptosis of photoreceptors in the ONL, and suppress the inflammatory response in the retina of RCS rats. In vitro, we have shown that hRPC-sEV treatment could significantly reserve the low-glucose preconditioned apoptosis of photoreceptors and reduce the expression of pro-inflammatory cytokines in microglia. Pathway analysis predicted the target genes of hRPC-sEV microRNAs involved in inflammation related biological processes and significantly enriched in processes autophagy, signal release, regulation of neuron death, and cell cycle. Collectively, our study suggests that hRPC-sEVs might be a favorable agent to delay retinal degeneration and highlights as a new paradigm for cell-free therapy.

scholarly journals Human embryonic stem cell-derived extracellular vesicles alleviate retinal degeneration by upregulating Oct4 to promote retinal Müller cell retrodifferentiation via HSP90

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Yifeng Ke ◽  
Xiaoe Fan ◽  
Rui Hao ◽  
Lijie Dong ◽  
Min Xue ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Extracellular Vesicles ◽  
Fluorescence Intensity ◽  
Embryonic Stem ◽  
Müller Cells ◽  
Muller Cells ◽  
Retinal Progenitor Cells ◽  
Retinal Progenitor ◽  
Hsp90 Expression ◽  
Rcs Rats

Abstract Objective Retinal degenerative diseases remain the dominant causes of blindness worldwide, and cell replacement is viewed as a promising therapeutic direction. However, the resources of seed cells are hard to obtain. To further explore this therapeutic approach, human embryonic stem extracellular vesicles (hESEVs) were extracted from human embryonic stem cells (hESCs) to inspect its effect and the possible mechanism on retinal Müller cells and retinal function. Methods hESEVs were extracted by multi-step differential centrifugation, whose morphologies and specific biomarkers (TSG101, CD9, CD63, and CD81) were observed and measured. After hESEVs were injected into the vitreous cavity of RCS rats, the retinal tissues and retinal functions of rats were assessed. The alteration of Müller cells and retinal progenitor cells was also recorded. Microvesicles (MVs) or exosomes (EXOs) were extracted from hESCs transfected with sh-HSP90 or pcDNA3.1-HSP9, and then incubated with Müller cells to measure the uptake of EVs, MVs, or EXOs in Müller cells by immunofluorescence. The retrodifferentiation of Müller cells was determined by measuring Vimentin and CHX10. qRT-PCR and western blot were used to detect HSP90 expression in MVs and evaluate Oct4 level in Müller cells, and Co-IP to inspect the interaction of HSP90 and Oct4. Results RCS rats at the postnatal 30 days had increased retinal progenitor cells which were dedifferentiated from Müller cells. hESEVs were successfully extracted from hESCs, evidenced by morphology observation and positive expressions of specific biomarkers (TSG101, CD9, CD63, and CD81). hESEVs promoted Müller cells dedifferentiated and retrodifferentiated into retinal progenitor cells evidenced by the existence of a large amount of CHX10-positive cells in the retinal inner layer of RCS rats in response to hESEV injection. The promotive role of hESEVs was exerted by MVs demonstrated by elevated fluorescence intensity of CHX10 and suppressed Vimentin fluorescence intensity in MVs rather than in EXOs. HSP90 in MVs inhibited the retrodifferentiation of Müller cells and suppressed the expression level of Oct4 in Müller cells. Co-IP revealed that HSP90 can target Oct4 in Müller cells. Conclusion hESEVs could promote the retrodifferentiation of Müller cells into retinal progenitor cells by regulating the expression of Oct4 in Müller cells by HSP90 mediation in MVs.

scholarly journals Subretinal Implantation of Electrospun, Short Nanowire, and Smooth Poly(ε-caprolactone) Scaffolds to the Subretinal Space of Porcine Eyes

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
A. T. Christiansen ◽  
S. L. Tao ◽  
M. Smith ◽  
G. E. Wnek ◽  
J. U. Prause ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Average Rate ◽  
Multifocal Electroretinography ◽  
Subretinal Space ◽  
Outer Retina ◽  
Retinal Progenitor Cells ◽  
Retinal Progenitor ◽  
Biodegradable Scaffolds ◽  
Porcine Eyes ◽  
Photoreceptor Damage

Biodegradable scaffolds play an important adjunct role in transplantation of retinal progenitor cells (RPCs) to the subretinal space. Poly(ε-Caprolactone) (PCL) scaffolds with different modifications were subretinally implanted in 28 porcine eyes and evaluated by multifocal electroretinography (mfERG) and histology after 6 weeks of observation. PCL Short Nanowire, PCL Electrospun, and PCL Smooth scaffolds were well tolerated in the subretinal space in pigs and caused no inflammation and limited tissue disruption. PCL Short Nanowire had an average rate of preserved overlying outer retina 17% higher than PCL Electrospun and 25% higher than PCL Smooth. Furthermore, PCL Short Nanowire was found to have the most suitable degree of stiffness for surgical delivery to the subretinal space. The membrane-induced photoreceptor damage could be shown on mfERG, but the reductions in P1 amplitude were only significant for the PCL Smooth. We conclude that of the tested scaffolds, PCL Short Nanowire is the best candidate for subretinal implantation.

Long-Term in vitro Expansion of Retinal Progenitor Cells by Culturing Intact Neurospheres in Monolayer

2008 ◽  
Vol 40 (6) ◽  
pp. 291-297 ◽  
Author(s):  
Pinghong Lai ◽  
Shibo Tang ◽  
Jinglin Yi ◽  
Xiaobo Zhu ◽  
Yuping Zou ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Retinal Progenitor Cells ◽  
Retinal Progenitor ◽  
In Vitro Expansion

scholarly journals Construction and Characterization of EGFP Reporter Plasmid Harboring Putative Human RAX Promoter for in vitro Monitoring of Retinal Progenitor Cells Identity

2020 ◽  
Author(s):  
Atefeh Atefi ◽  
Pendar Shojaei kojouri ◽  
Fereshteh Karamali ◽  
Shiva Irani ◽  
Mohammad Hossein Nasr Esfahani
Keyword(s):  
Progenitor Cells ◽  
Molecular Mechanisms ◽  
Embryonic Stem ◽  
Retinal Layer ◽  
Retinal Progenitor Cells ◽  
Regulatory Regions ◽  
Retina Development ◽  
Retinal Progenitor ◽  
Egfp Reporter

Abstract Background : In retinal degenerative disease, progressive and debilitating conditions result in deterioration of retinal cells and visual loss. In human, retina lacks the inherent capacity for regeneration. Therefore, regeneration of retinal layer from human retinal progenitor cells (hRPCs) is a challenging task and restricted in vitro maintenance of hRPCs remains as the main hurdle. Retina and anterior neural fold homeobox gene ( RAX ) play critical roles in developing retina and maintenance of hRPCs. In this study, for the first time regulatory regions of human RAX gene with potential promoter activity were experimentally investigated. Results: For this purpose, after in silico analysis of regulatory regions of human RAX gene, the expression of EGFP reporter derived by putative promoter sequences was first evaluated in 293T cells and then in hRPCS derived from human embryonic stem cells. The candidate region ( RAX -3258bp) showed the highest EGFP expression in hRPCs. This reporter construct can be used for invitro monitoring of hRPC identity and verification of an efficient culture medium for maintenance of these cells. Conclusions: Furthermore, our findings provide a platform for better insight into regulatory regions of human RAX gene and molecular mechanisms underlying its vital functions in retina development.

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