Perfusion Bioreactor Cell Banking for High-Density Viability
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Solution Overview
Problem
Current cell banking methods are inefficient for producing high-density cell cultures from frozen stocks due to low cell viability and the need for damaging concentration steps, limiting rapid production of high-density cultures.
Innovation Solution
The method involves culturing cells in a perfusion bioreactor with a cell retention system to achieve ultra-high density cell populations, which are then cryopreserved without additional concentration steps, maintaining high cell viability and allowing for direct production of ultra-high density frozen cell banks.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If cells are cryopreserved at low densities (1-2×10^7 cells/mL), then cell viability is maintained, but efficient inoculation of large volume cultures is not enabled
Solution Approach 1:
The patent changes the cell density parameter from conventional low density (1-2×10^7 cells/mL) to ultra-high density (≥1.0×10^8 cells/mL) by implementing perfusion culture technology. This parameter change enables both high productivity in inoculation and high cell density in the frozen stock, resolving the contradiction between these two requirements.
2Quantity of substance
If centrifugal concentration methods are used to create high density aliquots, then cell density is increased, but cell viability is reduced
Solution Approach 1:
The patent replaces the mechanical centrifugal concentration system with a perfusion culture system that maintains high cell density through continuous culture and selective retention. This substitution eliminates the damaging mechanical forces of centrifugation while achieving the same high density goal, thereby preserving cell viability.
Solution Approach 2:
The patent introduces a cell retention system as an intermediary component in the perfusion culture setup. This intermediary device enables high density cell accumulation without direct mechanical concentration, acting as a mediator that maintains both high cell density and high cell viability simultaneously.
3Productivity
If conventional cell banking methods are used, then existing protocols are maintained, but rapid production of high density cell cultures is not achieved
Solution Approach 1:
The patent implements continuous perfusion culture operation to maintain high cell density and viability throughout the culture process. This continuous operation enables rapid production of high density cell cultures by eliminating the interruptions and limitations of batch culture methods, significantly improving productivity.
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 approach enables the creation of ultra-high density cell banks with cell viability of at least 60% post-thaw, facilitating efficient production of high-density cell cultures without the need for centrifugal concentration, thereby improving the efficiency of cell banking processes.
Implementation Method 1
the cell retention system comprises an alternating tangential flow filtration system comprising a filter
Implementation Method 2
adding cryoprotectant to the ultra-high density cell population to produce an ultra-high density frozen cell bank
Data Source
AI summary
Provided are methods for the creation of ultra-high density cryopreserved cell banks. In certain embodiments, these methods employ altered perfusion culture techniques that allow for production of ultra-high density cell cultures that can be cryopreserved at unexpectedly high cell densities without the need for any cell concentration steps, while retaining excellent cell viability and quality.