A Scalable and Efficient Bioprocess for Manufacturing Human Pluripotent Stem Cell-Derived Endothelial Cells

Authors

Haishuang Lin, University of Nebraska - LincolnFollow
Qian Du, University of Nebraska - LincolnFollow
Qiang Li, University of Nebraska-LincolnFollow
Ou Wang, University of Nebraska - LincolnFollow
Zhanqi Wang, Beijing Anzhen Hospital of Capital Medical University
Neety Sahu, University of Nebraska-LincolnFollow
Christian Elowsky, University of Nebraska-LincolnFollow
Kan Liu, University of Nebraska-LincolnFollow
Chi Zhang, University of Nebraska-LincolnFollow
Soonkyu Chung, University of Nebraska-LincolnFollow
Bin Duan, University of Nebraska Medical CenterFollow
Yuguo Lei, University of Nebraska-LincolnFollow

Date of this Version

2018

Document Type

Article

Citation

Stem Cell Reports, Vol. 11, 454–469, August 14, 2018

Comments

Copyright 2018 The Authors.

Open access

https://doi.org/10.1016/j.stemcr.2018.07.001

Abstract

Endothelial cells (ECs) are of great value for cell therapy, tissue engineering, and drug discovery. Obtaining high-quantity and -quality ECs remains very challenging. Here, we report a method for the scalable manufacturing of ECs from human pluripotent stem cells (hPSCs). hPSCs are expanded and differentiated into ECs in a 3D thermoreversible PNIPAAm-PEG hydrogel. The hydrogel protects cells from hydrodynamic stresses in the culture vessel and prevents cells from excessive agglomeration, leading to high-culture efficiency including high-viability (>90%), high-purity (>80%), and high-volumetric yield (2.0 x 10⁷ cells/mL). These ECs (i.e., 3D-ECs) had similar properties as ECs made using 2D culture systems (i.e., 2D-ECs). Genome-wide gene expression analysis showed that 3D-ECs had higher expression of genes related to vasculature development, extracellular matrix, and glycolysis, while 2D-ECs had higher expression of genes related to cell proliferation.