Protocol for efficient generation of human artery and vein endothelial cells from pluripotent stem cells

Kyle M Loh; Sherry Li Zheng; Kevin J Liu; Qingqing Yin; Zhainib A Amir-Ugokwe; Sawan K Jha; Yue Qi; Vanessa K Wazny; Alana T Nguyen; Angela Chen; Faith-Masong Njunkeng; Christine Cheung; Edda Spiekerkoetter; Kristy Red-Horse; Lay Teng Ang

doi:10.1016/j.xpro.2024.103494

Protocol for efficient generation of human artery and vein endothelial cells from pluripotent stem cells

STAR Protoc. 2024 Dec 18;6(1):103494. doi: 10.1016/j.xpro.2024.103494. Online ahead of print.

Authors

Affiliations

¹ Institute for Stem Cell Biology & Regenerative Medicine, Stanford University, Stanford, CA 94305, USA; Department of Developmental Biology, Stanford University, Stanford, CA 94305, USA. Electronic address: [email protected].
² Institute for Stem Cell Biology & Regenerative Medicine, Stanford University, Stanford, CA 94305, USA; Department of Developmental Biology, Stanford University, Stanford, CA 94305, USA. Electronic address: [email protected].
³ Institute for Stem Cell Biology & Regenerative Medicine, Stanford University, Stanford, CA 94305, USA.
⁴ Institute for Stem Cell Biology & Regenerative Medicine, Stanford University, Stanford, CA 94305, USA; Department of Developmental Biology, Stanford University, Stanford, CA 94305, USA.
⁵ Institute for Stem Cell Biology & Regenerative Medicine, Stanford University, Stanford, CA 94305, USA; Howard Hughes Medical Institute, Department of Biology, Stanford University, Stanford, CA 94305, USA.
⁶ Howard Hughes Medical Institute, Department of Biology, Stanford University, Stanford, CA 94305, USA.
⁷ Division of Pulmonary, Allergy and Critical Care Medicine, Department of Medicine, Vera Moulton Wall Center for Pulmonary Vascular Disease, Stanford University, Stanford, CA 94305, USA.
⁸ Lee Kong Chian School of Medicine, Nanyang Technological University, 308232, Singapore, Singapore.
⁹ Institute for Stem Cell Biology & Regenerative Medicine, Stanford University, Stanford, CA 94305, USA; Department of Urology, Stanford University, Stanford, CA 94305, USA.
¹⁰ Lee Kong Chian School of Medicine, Nanyang Technological University, 308232, Singapore, Singapore; Institute of Molecular and Cell Biology, A∗STAR, 138673, Singapore, Singapore.
¹¹ Institute for Stem Cell Biology & Regenerative Medicine, Stanford University, Stanford, CA 94305, USA; Department of Urology, Stanford University, Stanford, CA 94305, USA. Electronic address: [email protected].

PMID: 39705146
DOI: 10.1016/j.xpro.2024.103494

Abstract

Blood vessels permeate all organs and execute myriad roles in health and disease. Here, we present a protocol to efficiently generate human artery and vein endothelial cells (ECs) from pluripotent stem cells within 3-4 days of differentiation. We delineate how to seed human pluripotent stem cells and sequentially differentiate them into primitive streak, lateral mesoderm, and either artery or vein ECs. We differentiate stem cells in defined, serum-free culture media in monolayers, without feeder cells or genetic manipulations. For complete details on the use and execution of this protocol, please refer to Ang et al. ¹.

Keywords: cell culture; cell differentiation; stem cells.