Somite Cell Differentiation Pathways from Pluripotent Stem Cells
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Solution Overview
Problem
Existing methods struggle to efficiently induce dermatome, syndetome, and mesenchymal stromal cells from pluripotent stem cells, and there is a lack of protocols for differentiating somite cells into these specific cell types and their derivatives.
Innovation Solution
A stepwise method involving the use of GSK3β inhibitors and BMP signaling to differentiate pluripotent stem cells into somite cells, which are then further differentiated into dermatome, myotome, sclerotome, and syndetome cells, and mesenchymal stromal cells through specific culture conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If Activin/Nodal/TGFβ signaling and BMP signaling are used to induce mesoderm from pluripotent stem cells, then mesoderm induction is achieved, but the induction mainly consists of lateral plate mesoderm rather than paraxial mesoderm
Solution Approach 1:
The patent changes the signaling parameters by blocking Activin/Nodal/TGFβ and BMP signaling pathways while activating WNT signaling, transforming the induction outcome from lateral plate mesoderm to paraxial mesoderm and subsequently to somite cells with high efficiency
2Productivity
If TGFβ inhibitor and high concentration GSK3 inhibitor are used to induce paraxial mesoderm, then induction rate reaches 70-95%, but the protocol complexity increases
Solution Approach 1:
The patent extracts and eliminates the need for TGFβ inhibitor by directly blocking Activin/Nodal signaling and using only GSK3 inhibitor to activate WNT signaling, simplifying the protocol while maintaining high induction rates of paraxial mesoderm and somite cells
3Adaptability or versatility
If somite cells are induced from pluripotent stem cells, then multiple differentiation capacity is achieved, but protocols for dermatome, syndetome, and mesenchymal stromal cells are lacking
Solution Approach 1:
The patent segments the differentiation process into distinct stages: first inducing somite cells from pluripotent stem cells via WNT activation, then separately inducing dermatome cells (via BMP inhibition), syndetome cells (via FGF and BMP), and mesenchymal stromal cells (via FGF), providing specific protocols for each cell type
Data Source
AI summary
Provided is a method of producing a somite cell from a pluripotent stem cell, comprising the step of culturing a pluripotent stem cell in a medium comprising a GSK3β inhibitor. Provided is a method of producing a dermatome cell from a somite cell, comprising the step of culturing a somite cell in a medium comprising a GSK3β inhibitor and BMP. Provided is a method of producing a syndetome cell from a sclerotome cell, comprising the steps of culturing a sclerotome cell in a medium comprising FGF and then culturing the cell in a medium comprising BMP and TGFβ. Provided is a method of producing a mesenchymal stromal cell from a somite cell, comprising the step of culturing a somite cell in a medium comprising FGF. Provided are methods of producing a myotome cell, a dermatome cell, a sclerotome cell, and a syndetome cell from a pluripotent stem cell through a somite cell by appropriately combining the above methods and known methods.