(176l) Manufacturing Human Pluripotent Stem Cell Derived Endothelial Cells in Scalable and Cell-Friendly Microenvironments

Wang, O., University of Nebraska-Lincoln
Lin, H., University of Nebraska, Lincoln
Li, Q., University of Nebraska, Lincoln
Du, Q., University of Nebraska-Lincoln
Zhang, C., University of Nebraska-Lincoln
Duan, B., University of Nebraska Medical Center
Lei, Y. L., University of Nebraska - Lincoln

Human pluripotent stem
cell derived endothelial cells (hPSC-ECs) are of great value for studying and
treating vascular diseases. However, manufacturing high quantity and quality
hPSC-ECs with current cell culture technologies remains very challenging. Here,
we report a novel method that can manufacture hPSC-ECs in scalable and
cell-friendly microenvironments to address this challenge. Using this method,
hPSCs are expanded and differentiated into ECs in microscale alginate hydrogel
tubes. The hydrogel tubes protect cells from the highly variable hydrodynamic
conditions and critical hydrodynamic stresses in the culture vessel and limit
the cell mass less than the diffusion limits (of human tissue) to ensure
efficient mass transport. The hydrogel tubes provide uniform and friendly
microenvironments for cells to grow. This novel design leads to extremely high
production efficiency. We showed that hPSC-ECs could be produced in 10 days
with high viability (>90%), high purity (>80%) and high yield (_5.0 ¡Á 108 cells per mL of microspace). The yield is
about 250 times that of the current-state-of-the-art. hPSC-ECs made in these
hydrogel tubes had similar in vitro and in vivo functions to hPSC-ECs generated
by conventional cell culture methods. This simple, scalable, efficient, defined
and cost-effective technology will make hPSC-ECs broadly available and
affordable for various biomedical applications.