Cytokine Culture for Isolating Gamma Delta T Cells
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Solution Overview
Problem
Current methods for isolating non-haematopoietic tissue-resident lymphocytes, particularly γδ T cells, face challenges in obtaining clinically relevant quantities due to significant cell loss and low yields, which limits their characterization and therapeutic applications.
Innovation Solution
A method involving culturing non-haematopoietic tissue samples in the presence of Interleukin-2 (IL-2), Interleukin-9 (IL-9), Interleukin-15 (IL-15), and Interleukin-21 (IL-21) to isolate and expand γδ T cells, maintaining tissue integrity and reducing inhibitory cell types, thereby increasing cell yield and purity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If conventional isolation methods are used to obtain non-haematopoietic tissue-resident lymphocytes, then the isolation process is simple, but the cell yield is low and clinically relevant quantities cannot be obtained
Solution Approach 1:
The patent applies preliminary action by culturing the tissue sample with cytokines (IL-2, IL-9, IL-15, IL-21) before isolation to pre-expand the γδ T cells. This pre-culturing step increases the cell yield before the actual isolation process, allowing clinically relevant quantities to be obtained without significantly increasing isolation complexity
Solution Approach 2:
The patent changes the chemical parameters of the culture environment by introducing specific cytokine combinations (IL-2, IL-9, IL-15, IL-21) to selectively expand γδ T cells. This parameter change enables high-yield isolation of clinically relevant cell quantities while maintaining a relatively simple isolation workflow
2Quantity of substance
If tissue samples are minced to isolate lymphocytes, then the isolation process is easier, but the structural integrity of the tissue is disrupted and cell yield is reduced
Solution Approach 1:
The patent performs preliminary cytokine culturing on intact or minimally processed tissue samples before isolation. This pre-expansion step increases γδ T cell numbers without requiring aggressive tissue disruption, thereby maintaining structural integrity while achieving high cell yields
Solution Approach 2:
The patent uses cytokine parameter changes (IL-2, IL-9, IL-15, IL-21) to selectively expand γδ T cells within the tissue structure. This chemical approach replaces mechanical disruption methods, preserving tissue integrity while achieving high cell recovery
3Quantity of substance
If conventional isolation methods are used, then the process is quick and simple, but the purity of γδ T cells is low due to presence of inhibitory cell types
Solution Approach 1:
The patent changes the biochemical parameters of the culture system by adding specific cytokine combinations (IL-2, IL-9, IL-15, IL-21) that selectively promote γδ T cell expansion while suppressing other cell types. This achieves high purity without requiring lengthy separation procedures
Solution Approach 2:
The patent converts the presence of multiple cell types in the tissue sample from a harmful factor into a benefit by using cytokine selectivity. The cytokine environment preferentially supports γδ T cell growth while limiting other cell types, effectively purifying the population without additional separation steps
Data Source
AI summary
The invention relates to a method for the isolation of lymphocytes (in particular γδ T cells) from a non-haematopoietic tissue sample comprising the steps of: culturing the non-haematopoietic tissue sample in the presence of (a) Interleukin-2 (IL-2) or Interleukin-9 (IL-9); (b) Interleukin-5 (IL-15); and (c) Interleukin-21 (IL-21); and collecting a population of lymphocytes cultured from the non-haematopoietic tissue sample. Methods of subsequent expansion are provided, as well as populations of isolated cells obtained by the method and uses thereof.