Non-expanded Multilineage Cell Isolation via Anti-SSEA-4 Antibody Sorting
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Solution Overview
Problem
Current methods for isolating and expanding post-natal multilineage-inducible cells for therapeutic use often result in cellular manipulation and contamination, requiring extensive testing and regulatory controls, and are unable to achieve sufficient cell quantities without culturing and expansion.
Innovation Solution
A method for isolating non-expanded post-natal multilineage-inducible cells from bone marrow using anti-SSEA-4 antibody conjugated with phycoerythrin, followed by magnetic or cell sorting, which allows for high purity and yield without expansion, maintaining the cells' multipotent characteristics.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If culturing and expansion procedures are used to increase cell quantities, then sufficient cell numbers for transplantation are achieved, but cellular manipulation and contamination occur requiring extensive testing and regulatory controls
Solution Approach 1:
The patent applies preliminary action by pre-coating culture dishes with extracellular matrix proteins (such as fibronectin, laminin, or collagen) before cell seeding. This preliminary preparation creates an optimized surface that enables cells to adhere and expand without requiring extensive manipulations or contaminations during the culturing process, thus achieving sufficient cell quantities while maintaining cell purity and safety
Solution Approach 2:
The patent uses extracellular matrix proteins as intermediary substances between the culture dish surface and the cells. These proteins mediate cell adhesion and proliferation, allowing cells to expand in a controlled manner without direct contact with potentially harmful culture conditions, thereby reducing contamination risks while achieving sufficient cell quantities for transplantation
2Quantity of substance
If culturing and expansion procedures are used to provide sufficient cell quantities, then therapeutic use is enabled, but cellular manipulation occurs altering cell structure requiring extensive testing
Solution Approach 1:
The patent applies preliminary action by pre-coating culture dishes with extracellular matrix proteins (such as fibronectin, laminin, or collagen) before cell seeding. This preliminary preparation creates an optimized surface that enables cells to adhere and expand without requiring extensive manipulations or contaminations during the culturing process, thus achieving sufficient cell quantities while maintaining cell purity and safety
Solution Approach 2:
The patent uses specific extracellular matrix proteins as culture substrates, which changes the physical and chemical parameters of the culture environment. This parameter change provides optimal conditions for cell adhesion and expansion that maintain cell structure integrity while enabling sufficient cell quantity production for therapeutic applications
3Quantity of substance
If traditional isolation methods are used, then cell quantities are obtained, but purity and yield are insufficient for direct therapeutic use
Solution Approach 1:
The patent replaces mechanical isolation methods with a biochemical approach using extracellular matrix protein coating. This substitution allows for selective adhesion and enrichment of target cells based on their surface properties, achieving both high yield and high purity (>90%) that enables direct therapeutic use without further purification steps
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 yields a high percentage of SSEA-4 positive cells, achieving over 3,500 times more cells than traditional methods, with greater than 98% purity and maintaining multipotent characteristics, enabling direct therapeutic use or later expansion.
Implementation Method 1
incubating the total nuclear cells in the presence of an antibody... using anti SSEA-4 antibody phycoerythrin conjugated SSEA-4-PE
Implementation Method 2
anti SSEA-4 antibody phycoerythrin conjugated SSEA-4-PE
Implementation Method 3
separating the cells to isolate non-expanded post-natal multilineage-inducible cells... magnetic or cell sorting
Data Source
Figure 1A~1D
Figure 2A~2D
AI summary
A method of isolating non-expanded post-natal multilineage-inducible cells has the steps of obtaining a biological sample from an animal; isolating bone marrow from the biological sample, isolating total nuclear cells (TNCs) obtained from the bone marrow, incubating the total nuclear cells in the presence of an antibody or a cell adhesion substrate, separating the cells to isolate non-expanded post-natal multilineage-inducible cells and isolating the non-expanded post-natal multilineage-inducible cells in the absence of expansion.