3D culturing of human pluripotent stem cells-derived endothelial cells for vascular regeneration

Edit Gara¹, Eleonora Zucchelli², Annamária Nemes¹, Zoltán Jakus^3,4, Kitti Ajtay^3,4, Éva Kemecsei^3,4, Gábor Kiszler⁵, Nikolett Hegedűs⁶, Krisztián Szigeti⁶, Iván Földes¹, Kristóf Árvai⁷, János Kósa⁷, Kraszimir Kolev⁸, Erzsébet Komorowicz⁸, Parasuraman Padmanabhan⁹, Pál Maurovich-Horvat¹, Edit Dósa¹, György Várady¹⁰, Miklós Pólos¹, István Hartyánszky¹, Sian E. Harding², Béla Merkely¹, Domokos Máthé⁶, Gábor Szabó^11,12, Tamás Radovits¹, Gábor Földes^1,2✉

1. Heart and Vascular Center, Semmelweis University, Budapest, H1122, Hungary.
2. National Heart and Lung Institute, Imperial College London, W12 0NN, United Kingdom.
3. Department of Physiology, Semmelweis University, Budapest, H1094, Hungary.
4. MTA-SE “Lendület” Lymphatic Physiology Research Group of the Hungarian Academy of Sciences and the Semmelweis University, Budapest, H1094, Hungary.
5. 3D HISTECH Ltd., Budapest, H1141, Hungary.
6. Department of Biophysics and Radiation Biology, Nanobiotechnology & In vivo Imaging Center, Semmelweis University, H1094, Budapest, Hungary and In vivo Imaging Advanced Core Facility, Hungarian Centre of Excellence for Molecular Medicine. www.hcemm.eu, Szeged, Hungary.
7. Department of Internal Medicine and Oncology, Semmelweis University; PentaCore Laboratory, Budapest, H1083, Hungary.
8. Department of Biochemistry, Institute of Biochemistry and Molecular Biology, Semmelweis University, Budapest, H1094, Hungary.
9. Lee Kong Chian School of Medicine, Imperial College - Nanyang Technological University, 636921, Singapore.
10. Research Centre for Natural Sciences, Budapest, H1117, Hungary.
11. Experimentelle Herzchirurgie, Ruprecht-Karls Universität, Heidelberg, 69120, Germany.
12. Department of Cardiac Surgery, University of Halle, Halle (Saale), 06108, Germany.

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Citation:
Gara E, Zucchelli E, Nemes A, Jakus Z, Ajtay K, Kemecsei É, Kiszler G, Hegedűs N, Szigeti K, Földes I, Árvai K, Kósa J, Kolev K, Komorowicz E, Padmanabhan P, Maurovich-Horvat P, Dósa E, Várady G, Pólos M, Hartyánszky I, Harding SE, Merkely B, Máthé D, Szabó G, Radovits T, Földes G. 3D culturing of human pluripotent stem cells-derived endothelial cells for vascular regeneration. Theranostics 2022; 12(10):4684-4702. doi:10.7150/thno.69938. Available from https://www.thno.org/v12p4684.htm

Abstract

Rationale: Human induced pluripotent stem cell-derived endothelial cells can be candidates for engineering therapeutic vascular grafts.

Methods: Here, we studied the role of three-dimensional culture on their characteristics and function both in vitro and in vivo.

Results: We found that differentiated hPSC-EC can re-populate decellularized biomatrices; they remain viable, undergo maturation and arterial/venous specification. Human PSC-EC develop antifibrotic, vasoactive and anti-inflammatory properties during recellularization. In vivo, a robust increase in perfusion was detected at the engraftment sites after subcutaneous implantation of an hPSC-EC-laden hydrogel in rats. Histology confirmed survival and formation of capillary-like structures, suggesting the incorporation of hPSC-EC into host microvasculature. In a canine model, hiPSC-EC-seeded onto decellularised vascular segments were functional as aortic grafts. Similarly, we showed the retention and maturation of hiPSC-EC and dynamic remodelling of the vessel wall with good maintenance of vascular patency.

Conclusions: A combination of hPSC-EC and biomatrices may be a promising approach to repair ischemic tissues.

Keywords: human pluripotent stem cells, endothelial cells, angiogenesis tracking, tissue-engineered vascular grafts, multimodality imaging