Porous-Membrane Cell Sheet Production for Easy Handling
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Solution Overview
Problem
Existing methods for producing cell sheets are complex and difficult to handle, and there is a need for a simpler and more efficient method that can be applied to a variety of cell types, including retinal pigment epithelial cells, mesenchymal stem cells, and fibroblasts, which addresses issues such as invasive surgery risks, inflammation, and shape changes during implantation.
Innovation Solution
A method involving culturing cells on a porous membrane in a cell culture vessel, where a porous membrane is used on the bottom, allowing for easy handling and separation of the cell sheet using physical forces like water flow, and includes the use of culture media with serum and/or Rho kinase inhibitors to enhance cell sheet formation and rigidity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If cells are cultured on artificial membrane or amniotic membrane, then cell sheet can be produced for implantation, but the composition and rigidity differ from basement membrane, inducing inflammation and rejection reaction
Solution Approach 1:
The invention allows cells to self-produce their own basement membrane through culture on a porous membrane. The cells differentiate and secrete extracellular matrix components, forming a basement membrane that is compositionally identical to in vivo basement membrane, thereby achieving excellent biocompatibility without inflammation or rejection reactions.
Solution Approach 2:
The invention changes the cultural parameters by using a porous membrane with specific pore size (0.1-10 μm) and surface properties that enable cell attachment and basement membrane formation. This parameter optimization allows cells to naturally produce the required extracellular matrix without artificial membrane interference.
2Ease of operation
If cell sheet is detached from planar substrate without special cultureware, then implantation procedure is simplified, but cell sheet shape changes easily before implantation
Solution Approach 1:
The invention uses a porous membrane that acts as a flexible support structure for the cell sheet. This membrane provides sufficient mechanical support to maintain cell sheet shape during handling and transport, yet allows for easy detachment and implantation without requiring special temperature-responsive cultureware or enzymatic treatment.
Solution Approach 2:
The basement membrane is formed on the side opposite to the substrate contact side during culture, creating a rigid structure that maintains cell sheet shape before implantation. This preliminary formation of structural support eliminates the need for complex handling procedures while preserving shape stability.
3Ease of manufacture
If cells are cultured without porous membrane, then culture method is simpler, but cell sheet cannot be easily separated and delivered to implantation site
Solution Approach 1:
The invention employs a porous membrane with optimized pore size (0.1-10 μm) that allows cell attachment and growth while enabling easy separation of the cell sheet from the membrane. The porous structure provides mechanical support during culture but permits simple detachment through water flow or gentle manipulation, facilitating easy delivery to implantation sites.
4Reliability
If complex culture methods are used to produce cell sheet, then cell sheet quality is improved, but production time and process complexity increase
Solution Approach 1:
The invention extracts the essential function of complex culture systems by using a simple porous membrane substrate that enables cells to naturally form basement membrane and maintain sheet structure. This extraction of the core supporting function eliminates the need for complex temperature-responsive cultureware, enzymatic treatment protocols, and multiple culture steps, significantly reducing production time while maintaining cell sheet quality.
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
The method enables easy handling and delivery of cell sheets to implantation sites without large incisions, maintaining their shape and rigidity, facilitating their use in medical applications.
Implementation Method 1
allowing for easy handling and separation of the cell sheet using physical forces like water flow
Implementation Method 2
includes the use of culture media with serum and/or Rho kinase inhibitors to enhance cell sheet formation and rigidity
Data Source
AI summary
The present invention aims to provide a simple method for producing a cell sheet, which allows easy handling of the cell sheet produced by the method. The present invention provides a method for producing a cell sheet, which includes: (1) a step of preparing one or more types of mammal-derived cells; (2) a step of seeding the prepared cells on a porous membrane of a first cell culture vessel having the porous membrane on its bottom; and (3) a step of placing the first cell culture vessel in a second cell culture vessel and culturing the cells, and which allows easy handling of the cell sheet produced by the method; and provides a cell sheet produced by the method.