MSC Purification by Centrifugal Filtration for Low BSA Aggregation
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Solution Overview
Problem
Existing methods for preparing mesenchymal stem cell (MSC) compositions result in high levels of xenogeneic substances, such as bovine serum albumin (BSA) and porcine trypsin, leading to allergic reactions, immune responses, and increased immunogenicity, while also causing MSC aggregation that can lead to pulmonary emboli, with current purification techniques failing to provide consistent and safe pharmaceutical compositions.
Innovation Solution
A method involving centrifugal filtration to reduce MSC aggregates to a size (D90 < 150 µm) and residual BSA to less than 55 µg/mL, using a closed system to minimize xenogeneic substances and maintain cell viability, thereby reducing the risk of allergic reactions and aggregation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional centrifugation methods are used to purify MSCs, then cell harvesting is achieved, but residual xenogeneic substances (BSA, porcine trypsin) remain causing immunogenicity and adverse reactions
Solution Approach 1:
The patent applies extraction by removing residual xenogeneic substances (BSA, porcine trypsin) from the MSC preparation through a multi-step purification process involving wash solutions and filtration, thereby eliminating the harmful factors while preserving the therapeutic cells
Solution Approach 2:
The patent changes the purification parameters by using specific wash solutions with controlled compositions and performing multiple washing cycles, which progressively reduces the concentration of xenogeneic contaminants to below detectable levels
2Object-affected harmful factors
If extensive washing is performed to reduce xenogeneic substances, then immunogenicity decreases, but cell viability and adhesive properties may be compromised
Solution Approach 1:
The patent uses wash solutions as intermediary substances that facilitate the removal of xenogeneic contaminants without directly contacting and damaging the MSCs. The wash solutions act as mediators that selectively remove harmful substances while preserving cell integrity
Solution Approach 2:
The patent performs multiple washing cycles (excessive action) to ensure complete removal of xenogeneic substances, but each individual wash is optimized to be gentle (partial action) on the cells, balancing thorough purification with cell preservation
3Productivity
If MSCs are expanded in media with FBS for cell growth, then cell expansion is achieved, but residual BSA accumulates causing adverse reactions including pulmonary embolism and anaphylaxis
Solution Approach 1:
The patent extracts residual BSA from the expanded MSC preparation through systematic washing and filtration processes, removing the harmful byproduct of cell expansion while preserving the expanded cell population for therapeutic use
Solution Approach 2:
The patent discards the wash solutions containing removed BSA and other contaminants, while recovering and preserving the MSCs that have been purified through the washing process, separating the harmful discarded material from the valuable recovered cells
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 reduces xenogeneic substances and MSC aggregation, enhancing the safety and efficacy of MSC compositions by minimizing allergic reactions and pulmonary emboli risk, with a 200-fold reduction in BSA and improved therapeutic outcomes.
Implementation Method 1
The cells are then washed by a centrifugal filtration process that simultaneously selects for cell mass and cell diameter
Implementation Method 2
The MSCs are then washed by a centrifugal filtration process
Data Source
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AI summary
One or more purified mesenchymal stem cell pharmaceutical compositions and methods of manufacture utilizing centrifugal filtration are disclosed. Threshold limits for intravenous administration of mesenchymal stem cell pharmaceutical compositions comprising residual animal products are also disclosed.