The convergence of cell biology and bioprocess engineering is creating fundamental new ways to impact disease. Advancements in human induced pluripotent stem cell (iPSC)-technologies have substantially expanded access to many human cell types to accommodate the future demand for such therapies. However, the direct utilization of standard cell manufacturing equipment and methods in the differentiation and manufacture of iPSC-derived cells can face significant challenges in obtaining the necessary production scales, quality standards and high reproducibility between batches for cost-effective cell therapy research and clinical application.
Currently, the development and production of iPSC-derived cell types is often performed in a small-scale culture, unsuitable for robust generation of a large number of cells. Stirred-tank bioreactors have emerged as promising culture systems for large-scale cell manufacturing from iPSC sources. These systems allow full automation and conduction in closed systems, resulting in cultures with comparable characteristics from batch to batch. Closed-system, parallel processing with increased automation is also critical to minimize error and contamination from human interaction with cell products.
Ncardia has established a controlled stirred-tank bioreactor platform that is shown to routinely yield high numbers of iPSC-derived endothelial cells and additional cell models. This scalable technology enables Ncardia to manufacture billions of high quality iPSC-derived cells, meeting an essential need for effective use in cell therapy, safety and efficacy applications, in terms of volume, safety and affordability. Using a Quality by Design approach, we demonstrate a robust and controlled process for large-scale manufacturing (>1x109) of iPSC-derived endothelial cells to a purity of >90% in a serum-free protocol.
The bioreactor-derived endothelial cells are shown to recapitulate angiogenesis in a capillary formation model, which is compatible with high-content imaging and high-throughput screening. Upon the formation of vascular lumen in microfluidic channels, sprouting into a three-dimensional collagen-based matrix was triggered with an optimized gradient of angiogenic factors.
We demonstrate the implementation of a flexible process development workflow comprised of state-of-the-art bioreactor systems that allows for optimization of processes at 15 mL scale, validation of promising conditions at pilot-scale (100 - 250 mL) and manufacture hiPSC-derived highly functional endothelial cells at a billion scale. This workflow, as part of our large-scale manufacturing platform, enables us to manufacture iPSC-derived cells of the highest quality and purity at any needed scale, meeting the needs of high cell-demanding therapies.