Cell Culture Medium for Stem Cell Derivation and Maintenance
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Solution Overview
Problem
Current cell culture protocols lack effective methods for obtaining sufficient quantities of stem cells from differentiated cells or maintaining them in an undifferentiated state, particularly for applications in cellular therapies and in vitro studies, such as for liver and pancreatic cells, due to the complexity of liver diseases and the need for alternative treatments to organ transplantation.
Innovation Solution
A cell culture medium comprising a base medium, an ABL and SRC dual kinase inhibitor, a mitogenic factor, and a WNT signaling pathway activator is used to prepare or maintain stem cells in an undifferentiated state, allowing for the derivation and stable passage of adult stem cells from various organs like the liver, pancreas, and colon, independent of FGF agonist, BMP antagonist, Notch agonist, and TGFβ inhibitor.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If conventional cell culture protocols are used, then existing cell culture methods can be maintained, but sufficient quantities of stem cells cannot be obtained from differentiated cells
Solution Approach 1:
The patent applies parameter changes by modifying the cell culture medium composition to include specific inhibitors (ABL and SRC kinase inhibitors) and activators (WNT pathway activator, mitogenic factor). This chemical parameter change enables differentiated cells to be converted into stem cells, thereby increasing the quantity of stem cells obtainable from differentiated cell sources.
2Stability of the object's composition
If conventional cell culture protocols are used, then existing methods can be maintained, but stem cells cannot be maintained in an undifferentiated state
Solution Approach 1:
The patent changes the chemical parameters of the culture medium by incorporating ABL and SRC kinase inhibitors along with WNT pathway activators. This compositional change stabilizes the undifferentiated state of stem cells during culture, preventing spontaneous differentiation while maintaining culture feasibility.
3Reliability
If organ transplantation is used to treat liver diseases, then liver function can be restored, but medical complexity increases and healthy liver graft availability is severely limited
Solution Approach 1:
The patent enables the generation of functional liver cells (hepatocytes) from stem cells that can be cultured in vitro. These cultured cells serve as a disposable alternative to whole organ transplantation, providing therapeutic effectiveness without the complexity and availability issues of organ transplantation. The stem cells can be expanded and differentiated as needed for cellular therapy.
4Reliability
If whole organ transplantation is used, then complete organ function can be restored, but severe lack of healthy organ availability occurs
Solution Approach 1:
The patent segments the whole organ transplantation approach into cellular-level therapy. Instead of transplanting entire organs, the invention enables transplantation of cultured stem cells or differentiated cells (such as hepatocytes or pancreatic beta cells) that can be mass-produced in vitro, thereby increasing availability while maintaining therapeutic reliability.
Data Source
AI summary
The present invention relates to a cell culture medium for preparing liver, gastric, pancreatic, colon or intestinal adult stem cell isolated from adult tissue, as well as for maintaining such stem cell in the undifferentiated state. The cell culture medium comprises a base medium; an ABL and SRC dual kinase inhibitor/an ABL kinase inhibitor and a SRC kinase inhibitor; a mitogenic factor; a WNT signalling pathway activator; a stimulator for NAD+ and NADP+ generation; and a cAMP/PKA pathway activator. In a particular embodiment, the ABL and SRC dual kinase inhibitor is Dasatinib; the mitogenic factor is EGF; the WNT signalling pathway activator is R-Spondin 1; the stimulator for NAD+ and NADP+ generation is nicotinamide; and the cAMP/PKA pathway activator is cholera endotoxin.