Hydrogel Cell Culture Detachment by Liquid Flow
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Solution Overview
Problem
Existing methods for detaching adherent cells from culture carriers using proteolytic enzymes can cause cell damage and contamination risks, and temperature-based detachment methods are inefficient for large-scale production, while polymer-based culture vessels are limited to planar cultures.
Innovation Solution
A method using a culture vessel with a hydrogel composed of a crosslinked vinyl alcohol-based polymer, where cells are cultured and detached using a low concentration of proteolytic enzyme and chelating agent, and detached via liquid flow, allowing for stable cell production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If proteolytic enzymes are used to detach cells from culture carriers, then cell detachment is achieved, but cell damage and contamination risks increase
Solution Approach 1:
The patent introduces a hydrogel as an intermediary substance between the culture carrier and cells. The hydrogel serves as a temporary scaffold that cells adhere to during culture, and can be easily removed without proteolytic enzymes, thus mediating the detachment process while protecting cells from enzyme damage and contamination
Solution Approach 2:
The patent replaces the chemical mechanism of proteolytic enzyme digestion with a physical mechanism of hydrogel removal. By using a soluble hydrogel that can be dissolved or washed away, the need for harsh chemical enzymes is eliminated, achieving cell detachment through physical means rather than chemical degradation
2Reliability
If temperature-based detachment methods are used, then cell detachment is achieved without enzymes, but production efficiency decreases for large-scale culture
Solution Approach 1:
The patent changes the key parameter from temperature to hydrogel solubility. Instead of using temperature changes that require prolonged cooling periods, the invention uses hydrogels with specific solubility characteristics that allow rapid removal, thus changing the detachment mechanism to achieve both safety and efficiency
3Reliability
If polymer-based culture vessels are used, then enzyme-free detachment is enabled, but planar culture limitation is imposed
Solution Approach 1:
The patent applies local quality by making only the surface layer of the culture carrier a hydrogel with specific solubility properties, while the bulk material can remain as a supportive polymer structure. This allows the surface to enable enzyme-free detachment while the internal structure provides mechanical support for three-dimensional cell growth
Solution Approach 2:
The patent uses composite materials by combining hydrogel with supportive polymer materials. The hydrogel provides the enzyme-free detachment capability on the surface, while the polymer matrix provides structural support, creating a composite culture carrier that enables both planar and three-dimensional cell cultures
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
Enables efficient and stable cell detachment without significant enzyme use, maintaining cell activity and suitability for large-scale production.
Implementation Method 1
a step of preparing a culture vessel comprising a composition comprising a liquid medium, a hydrogel containing a crosslinked body of a vinyl alcohol-based polymer, and raw material cells
Implementation Method 2
detaching the cultured cells from the surface of the hydrogel by bringing at least a part of the composition in the culture vessel into contact with a solution in which a concentration of a proteolytic enzyme is 1.5 mg/mL or less and a concentration of a chelating agent is 0.01 mg/mL or more
Data Source
AI summary
The present invention relates to a method for producing cells, comprising: a step of preparing a culture vessel comprising a composition comprising a liquid medium, a hydrogel containing a crosslinked body of a vinyl alcohol-based polymer, and raw material cells; a step of culturing the raw material cells on a surface of the hydrogel; and a step of detaching the cultured cells from the surface of the hydrogel by a liquid flow.