Rapid, efficient, and cost-effective cell programming platform
Current hiPSC differentiation protocols rely on an outside-in approach, where periodic application of external growth factors triggers intracellular pathways to control cellular differentiation. Growth factors are expensive. In addition, they lose bioactivity due to their fragile structures during cell culture. As cell differentiation is highly sensitive to the growth factor composition and concentration, the inevitable variation of growth factors leads to the production of mixed cell subtypes. It reduces the yield of producing desired cell types and increases the difficulty of cell purification. This project pioneers an intrinsic-in approach for rapid, cost-effective hiPSC programming by developing inducible transcription factor (TF) expression systems. This intrinsic-in approach leverages forward programming and forced TF expression to determine cell fate with high purity and speed. Additionally, this project will leverage state-of-the-art molecular engineering toolsets to target safe genomic sites with inducible expression to overcome bottlenecks in traditional protocols. Resultantly, an innovative approach will be developed to revolutionize large-scale hiPSC-derived beta cell production, paving the way for transformative therapeutic applications in diabetes treatment and beyond.
Scalable 3D microbioreactors for large-scale cell production
Current cell manufacturing infrastructure mainly depends on the use of conventional stirred tank bioreactors (STRs) for centralized cell production. This process of cell production is inherently associated with low cell densities, slow growth rates, and high cell damage or death during the production process. To satisfy the demand for producing a large number of cells, bioreactors are typically expanded in volume. However, simply increasing volume significantly increases cost and in turn brings scalability challenges due to inefficient production scales. Instead of merely scaling up bioreactor volume or quantity, we prioritize the development of a miniaturized cell culture system. A microbioreactor will be designed to mimic the three-dimensional stress-free tissue-like microenvironment found in vivo, overcoming the inefficiencies of traditional large-volume bioreactors. In a laboratory setting, we have already demonstrated groundbreaking efficiency, achieving 250 times increase in cell density compared to STRs, 3,000-fold cell expansion per passage, and reducing production costs by 90%. This significant leap in efficiency highlights the scalability and affordability of our microbioreactor, making it a potential game-changer for cell manufacturing. We will advance this microbioreactor design suitable for large-scale cell production and test the capability of new microbioreactors in producing hiPSC-derived beta cells.
Bioinspired equipment-free stress-free cell surface engineering
Transplanted cells face a survival challenge due to numerous stress factors such as the immune attack. Our lab has utilized DNA as a building block for cell surface engineering. The goal of this TR&D project is to finely tune this fundamental construction concept to meet the diverse requirements of cell-based applications across molecular, nanoscale, and microscale levels.