Bottom Support Liquid for Adherent Cell Tight Junction Formation
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Solution Overview
Problem
Current cell culture methods fail to effectively recreate the in vivo microenvironment, leading to changes in cell phenotypes and genotypes, and are limited by high costs and complex operations, particularly in forming tight junctions necessary for high cell density and accurate blood-brain barrier models.
Innovation Solution
A culture method involving adherent cells grown on a bottom support liquid with higher density than the culture medium, using fluorinated oils and siloxane compounds, promotes the formation of tight junctions and high cell density, enabling the creation of a membranous cell sheet suitable for 3D bioprinting and drug testing, and an in vitro blood-brain barrier model with a simple operation and low technical threshold.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If traditional solid matrix cell culture method is used, then the culture method is simple and low cost, but cell phenotypes and genotypes change and tight junction cannot be formed
Solution Approach 1:
The patent changes the physical parameter of the culture medium from conventional liquid to a density-modified liquid that is denser than the culture medium. This parameter change creates a unique interface environment that promotes tight junction formation and maintains cell phenotype stability while preserving the simplicity of the culture method.
Solution Approach 2:
The patent introduces an intermediary substance (the density-modified liquid) between the cells and the conventional liquid culture medium. This intermediary layer facilitates tight junction formation and maintains cell phenotype without requiring complex solid matrix structures or external forces.
2Stability of the object's composition
If three-dimensional solid matrix hydrogel is used, then suitable three-dimensional microenvironment is provided, but cell density is low and cost is high
Solution Approach 1:
The patent modifies the density parameter of the liquid culture medium to create a density gradient system. This parameter change enables cells to self-assemble into high-density structures with tight junctions while maintaining the simplicity of liquid culture, avoiding the need for expensive hydrogel matrices.
3Quantity of substance
If external force mediated solid matrix is used, then high cell density and tight junction are achieved, but operation is complex and learning threshold is high
Solution Approach 1:
The patent enables cells to self-assemble into high-density structures with tight junctions through the density-modified liquid environment. The cells automatically organize themselves without requiring external forces or complex operational procedures, making the method simple and easy to perform.
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 method allows for the formation of a membranous cell sheet with high cell density and tight junctions, enhancing the simulation of in vivo tissue structures, improving the accuracy and applicability of research in life science and clinical medicine, and facilitating the development of blood-brain barrier-related drugs with reduced operational complexity and cost.
Implementation Method 1
the bottom support liquid has higher density than the culture medium
Implementation Method 2
using fluorinated oils and siloxane compounds
Data Source
AI summary
The present disclosure provides a culture method of adherent cells generating a tight junction structure and a product thereof. The culture method includes: inoculating adherent cells resuspended in a culture medium onto a bottom support liquid in a culture container for culture to obtain a membranous cell sheet, where the bottom support liquid has higher density than the culture medium, and is not miscible with the culture medium. According to the culture method in the present disclosure, the bottom support liquid is applied to the culture of the adherent cells for the first time, the formation of a tight junction between the adherent cells and the secretion of a large number of extracellular matrices can be promoted so as to form a membranous cell sheet having high cell density, and in vivo tissue structures are well simulated.