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从健康和视网膜疾病特异性人诱导的多能干细胞中生成视网膜类器官

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Developmental Biology
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JoVE Journal Developmental Biology

Generation of Retinal Organoids from Healthy and Retinal Disease-Specific Human-Induced Pluripotent Stem Cells

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从健康和视网膜疾病特异性人诱导的多能干细胞中生成视网膜类器官

DOI:

10.3791/64509-v

•

09:47 min

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December 09, 2022

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Sudipta Mahato*^1,2, Trupti Agrawal*^1,2, Divya Pidishetty*^1,2, Savitri Maddileti, Vinay Kumar Pulimamidi³, Indumathi Mariappan

¹Centre for Ocular Regeneration, Prof. Brien Holden Eye Research Centre,LV Prasad Eye Institute, ²Manipal Academy of Higher Education, ³Schepens Eye Research Institute, Massachusetts Eye and Ear, Harvard Medical School

Chapters

00:05Introduction
01:19Passaging and Plating of hiPSCs to Initiate Retinal Lineage Differentiation
02:40Differentiation of hiPSCs into Eye Fields and Retinal Lineage
04:38Harvesting of Retinal Organoids
06:41Results: Characterization of Retinal Organoids and RPE Cells Derived From Normal and RB1^-/- Mutant iPSCs
09:06Conclusion

Summary

Automatic Translation

该协议描述了一种将hiPSCs分化为眼场簇并使用涉及贴壁和悬浮培养系统的简化培养条件生成神经视网膜类器官的有效方法。其他眼细胞类型，如RPE和角膜上皮，也可以在视网膜培养物中从成熟的眼视野中分离出来。

Tags

Keywords: Retinal Organoids Human Induced Pluripotent Stem Cells Retinal Degeneration Retinal Pigment Epithelium In Vitro Models Cell-based Therapies Eye Development Inherited Retinal Diseases Differentiation Adherent Culture Suspension Culture Retinal Cups Spatial Positioning Morphological Features Cell Dissociation IPSC Culture Differentiation Induction Medium BFGF Noggin

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