Mesh Pore Spheroid Culture for Uniform Formation and Counting
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Solution Overview
Problem
Conventional three-dimensional cell culture methods struggle to produce spheroids of uniform size and lack a convenient and accurate method for counting them.
Innovation Solution
A device comprising a substrate and a mesh member with protruding members, coated with hexanoyl glycol chitosan, which forms and counts spheroids by seeding cells in pores of specific dimensions, allowing for uniform size culture and easy recovery.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional three-dimensional cell culture methods (hanging-drop, rotary culture, centrifugal separation, micromolding) are used, then spheroids can be cultured, but the spheroids cannot be cultured in uniform size
Solution Approach 1:
The patent employs a mesh member with a regular array of pores as the core structure for spheroid culture. Each pore serves as a defined space that confines cells to form spheroids of uniform size. The pore dimensions (50-500 μm) directly control the spheroid size, ensuring manufacturing precision without complex culture procedures.
Solution Approach 2:
The culture device is segmented into multiple independent pores in a regular array pattern. Each pore acts as an independent culture chamber that physically separates cells, forcing them to form discrete spheroids of controlled size. This segmentation approach simplifies the culture method while achieving uniform spheroid production.
2Measurement precision
If conventional spheroid culture methods are used, then spheroids can be produced, but there is no convenient and accurate method for counting them
Solution Approach 1:
The mesh member with its regular array of pores serves a dual function: it both cultures the spheroids and provides the counting framework. Since each pore corresponds to one spheroid location, counting is simplified to counting the occupied pores in the regular array, eliminating the need for separate complex counting procedures.
Solution Approach 2:
The mesh member is designed to perform multiple functions simultaneously: it acts as both the culture substrate that forms uniform spheroids and the counting template that enables accurate enumeration. This multi-functionality resolves the contradiction between measurement precision and ease of operation.
3Productivity
If mass production of spheroids is required for Langerhans islet cell transplantation, then large quantities are needed, but conventional methods cannot produce uniform spheroids
Solution Approach 1:
The mesh member contains a regular array of numerous pores (e.g., 96, 144, or more depending on mesh size and pore spacing), enabling simultaneous culture of many spheroids in uniform sizes. This segmented approach allows mass production while maintaining size consistency, as each pore independently produces one uniform spheroid.
Solution Approach 2:
The pore size parameter of the mesh member (50-500 μm) can be selected based on the desired spheroid size and production scale. By changing the mesh specifications, the system can be optimized for different productivity requirements while maintaining uniform spheroid production, resolving the contradiction between quantity and consistency.
Data Source
AI summary
A device for forming and counting spheroids according to one embodiment of the present invention may comprise a substrate, and a mesh member which is configured to be able to be positioned on the surface of the substrate and has a plurality of protruding members. The plurality of protruding members intersects one another in the form of a mesh so as to form a plurality of pores.