Vascular Endothelial Cells Derived from Transgene-Free Pig Induced Pluripotent Stem Cells for Vascular Tissue Engineering

Luke Batty; Jinkyu Park; Lingfeng Qin; Muhammad Riaz; Yuyao Lin; Zhen Xu; Xuefei Gao; Xin Li; Colleen Lopez; Wei Zhang; Marie Hoareau; Meghan E Fallon; Yan Huang; Hangqi Luo; Jiesi Luo; Séverine Ménoret; Peining Li; Zhenting Jiang; Peter Smith; David H Sachs; George Tellides; Ignacio Anegon; Jordan S Pober; Pentao Liu; Yibing Qyang

doi:10.1016/j.actbio.2024.12.033

Vascular Endothelial Cells Derived from Transgene-Free Pig Induced Pluripotent Stem Cells for Vascular Tissue Engineering

Acta Biomater. 2024 Dec 14:S1742-7061(24)00743-8. doi: 10.1016/j.actbio.2024.12.033. Online ahead of print.

Authors

Luke Batty¹, Jinkyu Park², Lingfeng Qin³, Muhammad Riaz⁴, Yuyao Lin⁵, Zhen Xu⁴, Xuefei Gao⁶, Xin Li⁴, Colleen Lopez⁴, Wei Zhang⁴, Marie Hoareau⁴, Meghan E Fallon⁴, Yan Huang⁴, Hangqi Luo⁴, Jiesi Luo⁴, Séverine Ménoret⁷, Peining Li⁸, Zhenting Jiang⁹, Peter Smith¹⁰, David H Sachs¹¹, George Tellides³, Ignacio Anegon⁷, Jordan S Pober¹², Pentao Liu¹³, Yibing Qyang¹⁴

Affiliations

¹ Department of Internal Medicine, Section of Cardiovascular Medicine, Yale Cardiovascular Research Center, Yale School of Medicine, 300 George Street, New Haven, CT 06511, USA; Yale Stem Cell Center, 10 Amistad Street, New Haven, CT 06511, USA; Department of Pathology, Yale University, New Haven, CT 06510, USA.
² Department of Internal Medicine, Section of Cardiovascular Medicine, Yale Cardiovascular Research Center, Yale School of Medicine, 300 George Street, New Haven, CT 06511, USA; Yale Stem Cell Center, 10 Amistad Street, New Haven, CT 06511, USA; Department of Physiology, College of Medicine, Hallym University, Hallymdaehak-gil, Chuncheon-si, Gangwon-Do, 24252, South Korea.
³ Department of Surgery, Yale University, New Haven, CT 06520, USA.
⁴ Department of Internal Medicine, Section of Cardiovascular Medicine, Yale Cardiovascular Research Center, Yale School of Medicine, 300 George Street, New Haven, CT 06511, USA; Yale Stem Cell Center, 10 Amistad Street, New Haven, CT 06511, USA.
⁵ Department of Internal Medicine, Section of Cardiovascular Medicine, Yale Cardiovascular Research Center, Yale School of Medicine, 300 George Street, New Haven, CT 06511, USA; Yale Stem Cell Center, 10 Amistad Street, New Haven, CT 06511, USA; Department of Plastic, Aesthetic and Maxillofacial Surgery, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, 710061, China.
⁶ Department of Physiology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, China.
⁷ Center for Research in Transplantation and Translational Immunology UMR1064, INSERM, Nantes Université, Nantes, France; Nantes Université, SFR Santé, Inserm UMS 016, CNRS UMS 3556, Nantes, France.
⁸ Department of Genetics, Yale University, New Haven, CT 06519, USA.
⁹ Department of Earth & Planetary Sciences, Yale University, New Haven, CT 06511, USA.
¹⁰ Department of Comparative Medicine, Yale University, New Haven, CT 06520, USA.
¹¹ Department of Surgery, Columbia Center for Translational Immunology, Vagelos College of Physicians and Surgeons, Columbia University, New York, NY, USA; Department of Surgery, Massachusetts General Hospital, Boston, MA, USA.
¹² Department of Pathology, Yale University, New Haven, CT 06510, USA; Department of Immunobiology, Yale University, New Haven, CT 06520, USA.
¹³ School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Stem Cell and Regenerative Medicine Consortium, Pokfulam, Hong Kong.
¹⁴ Department of Internal Medicine, Section of Cardiovascular Medicine, Yale Cardiovascular Research Center, Yale School of Medicine, 300 George Street, New Haven, CT 06511, USA; Yale Stem Cell Center, 10 Amistad Street, New Haven, CT 06511, USA; Department of Pathology, Yale University, New Haven, CT 06510, USA; Department of Biomedical Engineering, Yale University, New Haven, CT 06519, USA. Electronic address: [email protected].

PMID: 39681154
DOI: 10.1016/j.actbio.2024.12.033

Abstract

Induced pluripotent stem cells (iPSCs) hold great promise for the treatment of cardiovascular diseases through cell-based therapies, but these therapies require extensive preclinical testing that is best done in species-in-species experiments. Pigs are a good large animal model for these tests due to the similarity of their cardiovascular system to humans. However, a lack of adequate pig iPSCs (piPSCs) that are analogous to human iPSCs has greatly limited the potential usefulness of this model system. Herein, transgene-free piPSCs with true pluripotency were generated by using reprogramming factors in an optimized pig pluripotency medium. Using an effective differentiation protocol, piPSCs were used to derive endothelial cells (ECs) which displayed EC markers and functionality comparable to native pig ECs. Further, piPSC-ECs demonstrated suitability for vascular tissue engineering, producing a tissue engineered vascular conduit (TEVC) that displayed the upregulation of flow responding markers. In an in vivo functional study, these piPSC-EC-TEVCs maintained the expression of endothelial markers and prevented thrombosis as interposition inferior vena cava grafts in immunodeficient rats. The piPSCs described in this study open up the possibility of unique preclinical species-in-species large animal modeling for the furtherance of modeling of cell-based cardiovascular tissue engineering therapies. STATEMENT OF SIGNIFICANCE: While there has been significant progress in the development of cellularized cardiovascular tissue engineered therapeutics using stem cells, few of them have moved into clinical trials. This is due to the lack of a robust preclinical large animal model to address the high safety and efficacy standards for transplanted therapeutics. In this study, pig stem cells that are analagous to human's were created to address this bottleneck. They demonstrated the ability to differentiate into functional endothelial cells and were able to create a tissue engineered therapeutic that is analogous to a human therapy. With these cells, future experiments testing the safety and efficacy of tissue engineered constructs are possible, bringing these crucial therapeutics closer to the patients that need them.

Keywords: Tissue engineering; endothelial cells; pig induced pluripotent stem cells; transgene-free.