Poster
2

Large-scale manufacture of hiPSC-derived endothelial cells for drug discovery and cell therapy

Authors

J Kamphorst1
1 Ncardia, Germany

Abstract

The convergence of cell biology and bioprocess engineering is creating fundamental new ways
to impact disease. Advancements in induced pluripotent stem
cell (hiPSC) 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 hiPSC-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 hiPSC 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 hiPSC-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. Total sprout area,
total sprout length and migration distance of each sprout were quantified using
high content imaging. We demonstrate that, similar to primary 

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