Amnion Stem Cell Isolation via Temperature-Controlled Enzymatic Digestion
Solution Overview
Problem
Current methods for isolating mesenchymal stem cells from amnion-derived tissues face challenges in removing epithelial cells, leading to impure cell populations due to the adherence nature of these cells, making it difficult to achieve high purity.
Innovation Solution
A method involving storing the amnion in a medium at controlled temperatures (−1° C. or higher and 25° C. or lower) for several hours, followed by enzymatic treatment with trypsin, collagenase, or dispase, and subsequent culturing to efficiently isolate mesenchymal stem cells while reducing epithelial cell inclusion.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If physical detaching of unnecessary tissue pieces is used to prevent epithelial cell inclusion, then some epithelial cells are removed, but epithelial cells existing inside the amnion cannot be removed
Solution Approach 1:
The patent applies preliminary action by storing the amnion in a medium at controlled temperatures (−1°C or higher and 25°C or lower) for several hours before enzymatic treatment. This pre-treatment step prepares the tissue for subsequent enzymatic digestion, making the separation of epithelial cells from mesenchymal stem cells more effective and easier to perform.
2Productivity
If enzymatic treatment is carried out immediately after cutting amnion into small pieces, then treatment efficiency is improved, but the ratio of epithelial cells in the cell population is significantly increased
Solution Approach 1:
The patent applies preliminary action by storing the amnion in a medium at controlled temperatures (−1°C or higher and 25°C or lower) for several hours before enzymatic treatment. This pre-treatment step prepares the tissue for subsequent enzymatic digestion, making the separation of epithelial cells from mesenchymal stem cells more effective and easier to perform.
Solution Approach 2:
The patent applies parameter changes by controlling the storage temperature of the amnion in the medium within a specific range (−1°C or higher and 25°C or lower) for several hours before enzymatic treatment. This temperature control modifies the physical state of the tissue, optimizing it for enzymatic digestion while minimizing epithelial cell inclusion in the final cell population.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach allows for the efficient isolation of mesenchymal stem cells with high purity, reducing the number of epithelial cells to a minimum, thereby facilitating the production of high-quality cell preparations at lower costs.
Implementation Method 1
storing the amnion in a medium at controlled temperatures (−1° C. or higher and 25° C. or lower) for several hours
Implementation Method 2
enzymatic treatment with trypsin, collagenase, or dispase
Data Source
AI summary
An object of the present invention is to provide a method of producing a cell population comprising mesenchymal stem cells, comprising efficiently isolating a cell population comprising mesenchymal stem cells at high purity from an amnion. Provided is a method of producing a cell population comprising amnion-derived mesenchymal stem cells, comprising (1) storing an amnion in a medium for 4 hours or longer at −1° C. or higher and 25° C. or lower, followed by (2) isolating the amnion from the medium and treating the amnion with an enzyme; and (3) culturing a cell fraction comprising mesenchymal stem cells after the treatment with the enzyme.