hEMP Cell Generation via Cluster Disruption for T Cell Therapy
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current T cell therapies rely on finite sources of human T cells, which limits their therapeutic potential, as they lack self-renewal capabilities, necessitating the development of efficient methods for generating T cells from stem cells.
Innovation Solution
The method involves generating human embryonic mesenchymal progenitor (hEMP) cells from non-clustered human embryonic stem (ES) or induced pluripotent stem (iPS) cells by seeding them at a defined single cell density on substrates like Matrigel or recombinant human vitronectin, cultivating them to a desired confluence, and then differentiating them into T cells using specific culture media and conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If T cells are derived from patient or normal donor, then they can target and kill cancer cells, but they lack self-renewal and are finite in supply
Solution Approach 1:
The patent applies preliminary action by establishing a stem cell bank before therapeutic need arises. Pluripotent stem cells are harvested, cryopreserved, and stored in advance, allowing unlimited future expansion into T cells when needed, thus resolving the contradiction between finite donor supply and unlimited therapeutic demand
Solution Approach 2:
The patent employs self-service through the self-renewal capability of pluripotent stem cells. These cells can indefinitely self-renew in culture and differentiate into T cells on demand, providing an autonomous, unlimited source of therapeutic cells without requiring repeated donations from patients or donors
2Productivity
If stem cells are cultivated at high density, then cell growth is accelerated, but differentiation efficiency decreases
Solution Approach 1:
The patent applies dynamics by implementing dynamic control of cell density throughout the differentiation process. The method uses a multi-stage approach where cell density is optimized at each phase: initial expansion at higher density followed by dilution to lower density during differentiation induction, allowing both rapid growth and high differentiation efficiency to be achieved at different times
Solution Approach 2:
The patent uses preliminary action by pre-expanding stem cells to sufficient numbers before initiating differentiation. This allows the culture to reach an optimal cell density threshold that ensures robust differentiation when induction factors are added, preventing differentiation failure while maintaining overall process efficiency
3Reliability
If clusters of stem cells are used, then cell survival is improved, but differentiation into single-cell-derived T cells is compromised
Solution Approach 1:
The patent applies preliminary action by performing mechanical or enzymatic disruption of cell clusters before differentiation induction. This preliminary step breaks up aggregates into single cells while maintaining cell viability through optimized disruption conditions, ensuring that subsequent T cell derivation comes from truly clonal single-cell origins
Solution Approach 2:
The patent uses an intermediary approach by introducing a cluster disruption step as a bridge between cluster formation and single-cell differentiation. This intermediate treatment phase uses gentle mechanical agitation or enzymatic agents to separate clusters without damaging cells, then allows cells to reattach as single cells before differentiation begins
Data Source
AI summary
The disclosure provides a method of generating non-clustered stem cells. Cluster disruption prior to mesoderm differentiation increases yield and efficiency in hEMP and T cell differentiation. Thus, this method allows the development of improved methods of hEMP and T cell differentiation.