Genetic Circuit Sequencing for Hematopoietic Cell Programming
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Solution Overview
Problem
Existing methods for converting stem cells to hematopoietic lineage cells are inefficient and require exogenous factors like thrombopoietin (TPO), FLT-3 Ligand (FLT3L), and interleukin (IL), which can complicate the process and reduce conversion rates.
Innovation Solution
A method involving a heterologous genetic circuit with gate units that modulate expression levels of distinct target genes sequentially, without exogenous factors, to convert stem cells to hematopoietic lineage cells, using a medium free of TPO, FLT3L, and IL, achieving a conversion rate of at least 5% within 14 days.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If exogenous factors (TPO, FLT3L, IL) are used in stem cell culture medium, then cell survival and proliferation are supported, but conversion rate to hematopoietic lineage cells is reduced and process complexity increases
Solution Approach 1:
The patent removes exogenous factors (TPO, FLT3L, IL) from the culture medium formulation, extracting the problematic elements that were hindering conversion efficiency. This extraction allows the stem cells to differentiate into hematopoietic lineage cells more effectively without the interfering effects of these exogenous growth factors, thereby improving conversion rate while simplifying the culture system.
Solution Approach 2:
The patent enables the stem cells to utilize their endogenous factors and intrinsic differentiation pathways rather than relying on externally added exogenous factors. The cells are provided with a defined medium that supports their natural differentiation process, allowing them to self-regulate and convert to hematopoietic lineage cells with higher efficiency and specificity.
2Productivity
If exogenous factors (TPO, FLT3L, IL) are included in culture medium, then cell growth is maintained, but conversion rate to CD45+ hematopoietic lineage cells decreases
Solution Approach 1:
The patent extracts and eliminates exogenous factors (TPO, FLT3L, IL) from the culture medium that were previously maintaining cell growth but hindering conversion. By removing these factors, the system achieves higher conversion rates to CD45+ hematopoietic lineage cells while maintaining reliable cell growth through the defined medium composition and endogenous cell factors.
Solution Approach 2:
The patent changes the chemical composition parameters of the culture medium by formulating a defined medium without exogenous growth factors. This parameter change fundamentally alters the differentiation trajectory of stem cells, promoting efficient conversion to hematopoietic lineage cells while maintaining growth through adjusted medium components and endogenous regulation.
3Productivity
If conventional culture methods are used, then process simplicity is maintained, but conversion rate to hematopoietic lineage cells is low
Solution Approach 1:
The patent employs a defined medium that supports stem cell differentiation through endogenous factors and intrinsic pathways rather than requiring complex additions of exogenous growth factors. This self-service approach simplifies the manufacturing process while achieving high conversion rates to hematopoietic lineage cells, as the medium formulation is streamlined and does not require precise control of multiple exogenous components.
Data Source
AI summary
Provided herein are systems of modulating gene expression, methods of use thereof, and cells engineered thereof for the purpose of differentiating cells, for example hematopoietic cells.