Corneal Endothelial Cell Molecular Markers for Isolation
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Solution Overview
Problem
Current methods for isolating and evaluating corneal endothelial cells are inadequate due to the lack of specific molecular markers, leading to inefficiencies in corneal transplantation and treatment of corneal endothelial dysfunction.
Innovation Solution
Identification and utilization of specific genes such as ZP4, MRGPRX3, GRIP1, GLP1R, HTR1D, and CLRN1 as molecular markers to sort and detect corneal endothelial cells, enabling efficient isolation and evaluation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If current non-specific markers (ZO-1, Na+-K+-ATPase, N-cadherin) are used for corneal endothelial cell isolation, then cell detection can be performed, but the isolation precision is insufficient due to non-specific expression in many other cell types
Solution Approach 1:
The patent uses molecular markers (specific genes or proteins) as unique identifiers or 'signatures' for corneal endothelial cells. By detecting the presence and expression levels of these specific markers, the invention enables precise identification and isolation of corneal endothelial cells from mixed cell populations, resolving the contradiction between detection capability and isolation precision.
2Quantity of substance
If stem cells are used to induce corneal endothelial cells, then cell supply can be increased, but the cell population contains different types of cells requiring specific marker-based sorting
Solution Approach 1:
The patent employs specific molecular markers as intermediaries to bridge the gap between heterogeneous stem cell populations and pure corneal endothelial cell populations. These markers serve as mediators that enable selective identification and isolation of the desired cell type through techniques such as flow cytometry or magnetic-activated cell sorting, thereby simplifying the cell sorting process while maintaining high purity.
3Quantity of substance
If repeated passages are performed to expand corneal endothelial cells, then cell quantity increases, but genetic transformation occurs leading to loss of cell functions
Solution Approach 1:
The patent applies molecular marker-based sorting at an early stage to obtain a pure population of corneal endothelial cells before extensive culture passages are performed. By establishing high-purity cell populations upfront using specific markers, the invention prevents the accumulation of genetic transformations and functional losses that would otherwise occur during repeated passages, thereby maintaining cell reliability while achieving sufficient cell quantity.
Data Source
AI summary
A molecular marker expressed specifically in corneal endothelial cells, and a method for producing one or more corneal endothelial cells using the marker and a method for evaluating one or more corneal endothelial cells using the marker are provided. At least one molecule selected from the group consisting of ZP4, MRGPRX3, GRIP1, GLP1R, HTR1D, and CLRN1 is used as a marker specific to corneal endothelial cells.