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

Haishuang Lin; Qian Du; Qiang Li; Ou Wang; Zhanqi Wang; Neety Sahu; Christian Elowsky; Kan Liu; Chi Zhang; Soonkyu Chung; Bin Duan; Yuguo Lei

doi:10.1016/j.stemcr.2018.07.001

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

Stem Cell Reports. 2018 Aug 14;11(2):454-469. doi: 10.1016/j.stemcr.2018.07.001. Epub 2018 Aug 2.

Authors

Haishuang Lin¹, Qian Du², Qiang Li³, Ou Wang³, Zhanqi Wang⁴, Neety Sahu¹, Christian Elowsky⁵, Kan Liu², Chi Zhang², Soonkyu Chung⁶, Bin Duan⁷, Yuguo Lei⁸

Affiliations

¹ Department of Chemical and Biomolecular Engineering, University of Nebraska-Lincoln, Lincoln, NE 68588, USA.
² Department of Biological Systems Engineering, University of Nebraska-Lincoln, Lincoln, NE 68588, USA.
³ Department of Chemical and Biomolecular Engineering, University of Nebraska-Lincoln, Lincoln, NE 68588, USA; Biomedical Engineering Program, University of Nebraska-Lincoln, Lincoln, NE 68588, USA.
⁴ Department of Vascular Surgery, Beijing Anzhen Hospital of Capital Medical University, Beijing Institute of Heart Lung and Blood Vessel Diseases, Beijing 100029, China.
⁵ Department of Agronomy and Horticulture, University of Nebraska-Lincoln, Lincoln, NE 68588, USA.
⁶ Department of Nutrition and Health Sciences, University of Nebraska-Lincoln, Lincoln, NE 68583, USA.
⁷ Mary and Dick Holland Regenerative Medicine Program, University of Nebraska Medical Center, Omaha, NE 68198, USA.
⁸ Department of Chemical and Biomolecular Engineering, University of Nebraska-Lincoln, Lincoln, NE 68588, USA; Biomedical Engineering Program, University of Nebraska-Lincoln, Lincoln, NE 68588, USA; Mary and Dick Holland Regenerative Medicine Program, University of Nebraska Medical Center, Omaha, NE 68198, USA; Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center, Omaha, NE 68198, USA. Electronic address: ylei14@unl.edu.

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 × 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.

Keywords: 3D culture; endothelial cells; human pluripotent stem cells; thermoreversible hydrogel.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Batch Cell Culture Techniques*
Biomarkers
Bioreactors*
Cell Culture Techniques
Cell Differentiation
Computational Biology / methods
Endothelial Cells / cytology*
Endothelial Cells / metabolism*
Gene Expression Profiling
Gene Ontology
Humans
Hydrogels
Immunophenotyping
Pluripotent Stem Cells / cytology*
Pluripotent Stem Cells / metabolism*
Transcriptome

Substances

Biomarkers
Hydrogels