Ciliary Marginal Zone-Like Structure Formation for Pure Neural Retina
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Solution Overview
Problem
There is no known method for producing a ciliary marginal zone-like structure from pluripotent stem cells with high efficiency.
Innovation Solution
A method involving the formation of a cell aggregate comprising a ciliary marginal zone-like structure, retinal pigment epithelium, and neural retina, where the neural retina is present on the surface and forms a continuous, stratified layer, allowing for the production of a highly pure neural retina by cutting it out from the aggregate.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional methods are used to produce ciliary marginal zone-like structure from pluripotent stem cells, then the structure can be formed, but the production efficiency is low
Solution Approach 1:
The invention applies preliminary action by pre-forming cell aggregates with specific compositions containing pluripotent stem cells, Fezf2-expressing cells, and Chx10-expressing cells before initiating the differentiation process. This pre-organization of cell types and their spatial arrangement accelerates subsequent development into ciliary marginal zone-like structures, reducing overall production time while maintaining high efficiency
2Manufacturing precision
If the neural retina is present on the surface of the cell aggregate, then it forms a continuous stratified layer, but the complexity of isolating pure neural retina increases
Solution Approach 1:
The invention applies the extraction principle by isolating the neural retina from the surface of the cell aggregate through a simple mechanical cutting or detachment process. The neural retina, being positioned on the surface as a continuous stratified layer, can be easily separated from the underlying cell aggregate, yielding highly pure neural retina tissue without requiring complex isolation procedures
Data Source
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AI summary
The present invention provides a method for producing a cell aggregate containing a ciliary marginal zone-like structure, including a step of culturing a cell aggregate containing a retinal tissue in which Chx10 positive cells are present in a proportion of 20% or more of the tissue in a serum-free medium or serum-containing medium each containing a substance acting on the Wnt signal pathway for only a period before the appearance of a RPE65 gene expressing cell, followed by culturing the "cell aggregate in which a RPE65 gene expressing cell does not appear" thus obtained in a serum-free medium or serum-containing medium each not containing a substance acting on the Wnt signal pathway and so on. According to the present invention, a ciliary marginal zone-like structure can be produced with high efficiency.