Enzymatic Cell Separation in Biochamber for Viability
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Solution Overview
Problem
Current methods for separating cells from culture components in cell therapy suffer from cellular damage due to mechanical forces, leading to reduced viability and functionality, and significant loss of cells during the separation process, especially for mixed cell populations.
Innovation Solution
A cell washing procedure that minimizes exposure to mechanical forces, using a biochamber where a biocompatible rinse solution and enzyme solution are used to dissociate cells, followed by a second rinse solution to collect cells with minimal residual culture components, ensuring high viability and purity for clinical use.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional mechanical separation methods are used to separate cells from culture components, then cell separation efficiency is improved, but cell viability and functionality deteriorate due to mechanical damage
Solution Approach 1:
The patent replaces mechanical separation methods with a chemical-biological approach using enzymes (trypsin, collagenase) to dissociate cells from culture surfaces and components. This enzymatic digestion method eliminates mechanical stress on cells while achieving effective separation, thereby maintaining cell viability and functionality throughout the processing procedure.
Solution Approach 2:
The patent introduces enzyme solutions as intermediary agents between the cells and culture components. These enzymes act as mediators that selectively break down protein bonds holding cells to the culture surface without directly contacting or damaging the cells themselves, enabling separation while preserving cell integrity.
2Ease of manufacture
If conventional separation procedures are used, then cell processing is completed, but significant cell loss occurs during the separation process
Solution Approach 1:
The patent performs preliminary enzymatic treatment of the culture surface before cell detachment. By pre-incubating the culture surface with enzymes to weaken adhesion bonds, the cells can be gently released with minimal mechanical force, significantly reducing cell loss during the separation process while ensuring complete processing.
Solution Approach 2:
The patent changes the chemical parameters of the separation process by using controlled enzymatic reactions instead of mechanical forces. By adjusting enzyme concentration, incubation time, and temperature, the process achieves optimal cell release with minimal loss, transforming a mechanically aggressive process into a chemically controlled gentle separation.
3Quantity of substance
If cell culture steps are performed to increase cell number and purity, then cell quantity and purity are improved, but cell function is lost due to cell aging and contamination risk increases
Solution Approach 1:
The patent performs preliminary expansion of stem cells in culture before the separation procedure. By pre-culturing cells to achieve the desired quantity and purity, then immediately proceeding to enzymatic separation and direct transplantation, the method minimizes the time cells spend in culture post-expansion, thereby preserving cell function while achieving the necessary cell numbers.
Solution Approach 2:
The patent establishes a continuous process flow where cell expansion, enzymatic separation, and transplantation occur in sequence without interruption. This continuous action minimizes the time cells are exposed to culture conditions that could lead to aging or contamination, maintaining cell functionality throughout the process while achieving required cell quantities.
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
The method significantly enhances cell viability and yield, reducing residual culture components and microbial contamination, resulting in a cell product suitable for direct patient administration with improved shelf-life and cryopreservation potential.
Implementation Method 1
incubating the contents of the biochamber for a predetermined period of time with the enzyme solution such that during incubation, the enzyme at least dissociates the cells from each other and/or from the biochamber surface
Data Source
AI summary
The present invention provides a fluid exchange cell culture technique and tissue repair cells (TRCs) made by these methods, as well as methods using these cells. The method includes a new wash step which increases the tissue repair properties of the TRCs of the invention. This wash step allows for the production of TRC populations with greater tissue repair and anti-inflammatory capabilities. Embodiments of the present invention include a post-culture process for cultured cells that preferably includes the steps of: a wash process for removing unwanted residual culture components, a volume reduction process, and a harvesting process to remove cultured cells. Preferably, all these steps are performed within a aseptically closed cell culture chamber by implementing a separation method that minimizes mechanical disruption of the cells and is simple to automate. The harvested cells may then be concentrated to a final volume for the intended use. In such embodiments, the final composition is a substantially purified and concentrated cell mixture suspended in a physiologic solution suitable for immediate use in humans without further washing, volume reduction, or processing. Embodiments are also applicable to harvesting (and/or washing) particles within a liquid or solution within a chamber.