Single-Vessel Adherent Cell Amplification Using Dynamic Microcarriers
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Solution Overview
Problem
Current methods for large-scale production of adherent cells and biological agents are costly due to the need for multiple bioreactors, enzymatic treatments, and cell passage heterogeneity, which can lead to cell aging and reduced regulatory compliance.
Innovation Solution
A process where adherent cells are amplified in a single culture container through successive cell passages using enzymatic treatment to detach cells from microcarriers, with increasing microcarrier concentrations and medium volumes, reducing the need for multiple bioreactors and maintaining cell homogeneity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If multiple bioreactors are used for successive cell passages, then cell production scale is increased, but equipment complexity and production costs increase
Solution Approach 1:
The patent merges multiple cell passage operations into a single bioreactor system. The bioreactor is designed to accommodate sequential addition of microcarriers and cells, allowing multiple passages to occur in one vessel rather than requiring transfer between multiple bioreactors. This reduces equipment complexity while maintaining productivity.
Solution Approach 2:
The cell culture process is segmented into discrete stages within the same bioreactor: initial seeding, first passage completion, enzymatic treatment, microcarrier removal, and sequential addition of fresh microcarriers for subsequent passages. This segmentation allows complex multi-passage protocols to be executed in a single vessel through structured process steps.
2Productivity
If cells are transferred between multiple bioreactors, then cell amplification is achieved, but cell homogeneity is reduced due to passage heterogeneity
Solution Approach 1:
By combining all cell passage operations within a single bioreactor, the patent ensures that all cell populations undergo the same number of passages under identical conditions. This eliminates the heterogeneity that arises when cells are distributed across multiple bioreactors with different passage histories, thereby maintaining cell homogeneity while achieving amplification.
3Productivity
If enzymatic treatment is used to detach cells from microcarriers, then cell transfer efficiency is improved, but cell integrity is compromised
Solution Approach 1:
The patent extracts cells from microcarriers using brief enzymatic treatment with trypsin, then immediately removes the enzyme by centrifugation and washing with fresh medium. This minimizes the exposure time of cells to the harmful enzymatic action while still achieving efficient detachment, thus balancing transfer efficiency with cell integrity preservation.
Solution Approach 2:
The enzymatic treatment step is executed quickly and efficiently, then the process rushes through to cell washing and resuspension. The brief exposure to trypsin is sufficient for detachment but minimized to reduce damage, and the rapid subsequent washing removes the harmful enzyme before it can cause significant cell degradation.
4Productivity
If microcarrier concentration is increased to boost cell production, then cell biomass increases, but mass transfer limitations and cell stress increase
Solution Approach 1:
The patent dynamically adjusts microcarrier concentration throughout the cell culture process. Rather than maintaining a constantly high concentration, the system starts with lower concentrations during early passages when cell biomass is low, then progressively increases microcarrier concentrations in subsequent passages as cell populations grow. This dynamic adjustment optimizes cell production while preventing excessive cell stress from overcrowding and mass transfer limitations.
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 significantly reduces production costs and maintains cell homogeneity, allowing for the production of adherent cells and biological agents with enhanced efficiency and regulatory compliance by minimizing equipment use and cell passage variations.
Implementation Method 1
each cell passage subsequent to the first cell passage is carried out by using all or part of the cell population which was obtained during the previous cell passage after subjecting the cell population to enzymatic treatment to detach the cells from the microcarriers
Data Source
Figure 1a~1b
Figure 2a~2b
AI summary
The present invention relates to a method for producing adherent cells according to which: a. adherent cells are injected into a culture container which contains microcarriers in a culture medium; b. the cells are amplified by performing a plurality of consecutive cell migrations in said same culture container, wherein each cell migration after the first cell migration is carried out: i. using all or part of the cell population obtained during the preceding cell migration, after having subjected said cell population to an enzymatic treatment in order to separate the cells from the microcarriers, and ii. by injecting culture medium and an increasing amount of microcarriers; and c. the cell population produced during the last cell migration is collected after having optionally subjected said cell population to an enzymatic treatment in order to separate the cells from the microcarriers. The invention also relates to the implementation of said method for the production of biological agents, in particular for preparing vaccines or drugs.