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Identifying an optimal seeding protocol and endothelial cell substrate for biohybrid lung development

journal contribution
posted on 01.03.2019 by Ulrich Zwirner, Klaus Hoffler, Michael Pflaum, Sotiris Korossis, Axel Haverich, Bettina Wiegmann
© 2018 John Wiley & Sons, Ltd. Several key prerequisites need to be fulfilled for the development of a biohybrid lung, which can offer an actual alternative to lung transplantation. A major aspect is an optimized haemocompatibility of the device's artificial surfaces via endothelial cell seeding. In this study, four different types of polymeric gas exchange hollow fibre membranes (HFMs) were analysed utilizing four different seeding protocols in order to identify the ideal combination for sufficient long-term endothelialization. Human cord blood-derived endothelial cells (HCBECs) were used for the endothelialization of polypropylene HFMs with two different pore sizes and poly-4-methyl-1-pentene HFMs, both with and without heparin/albumin coating. The qualitative and quantitative impact of four different rotational seeding protocols regarding long-term HFM endothelialization and the impact of inflammatory stimulation on the seeded HCBECs were examined by fluorescence microscopy, cell counting, and analysis of relative expression levels of activation, shear stress, and thrombogenic state markers. Optimized endothelial cell seeding and long-term cultivation were only achieved using heparin/albumin-coated poly-4-methyl-1-pentene HFMs, applying 24 hr of rotational speed at 1 rpm followed by 120 hr of static culture. Neither cell-to-HFM contact nor the rotational cultivation procedure showed an impact on the physiological anti-thrombogenic and anti-inflammatory HCBEC activation status. Additionally, the cells maintained their physiological responsiveness towards inflammatory stimulation. Rotational seeding strategies and a seamless heparin/albumin coating of the HFMs are crucial requirements for a sufficient and long-lasting endothelialization and thus a key element in the future development and in vivo application of the biohybrid lung.

Funding

German Research Foundation (DFG), Grant/ Award Number: FI 1776/4‐2; German Center for Lung Research (DZL), Grant/Award Number: 82DZL00201; REBIRTH ‐ Cluster of Excellence, Grant/Award Number: EXC 62/1 (Unit 4.1)

History

School

  • Mechanical, Electrical and Manufacturing Engineering

Published in

Journal of Tissue Engineering and Regenerative Medicine

Volume

12

Issue

12

Pages

2319 - 2330

Citation

ZWIRNER, U. ... et al., 2018. Identifying an optimal seeding protocol and endothelial cell substrate for biohybrid lung development. Journal of Tissue Engineering and Regenerative Medicine, 12(12), pp. 2319 - 2330.

Publisher

© Wiley

Version

VoR (Version of Record)

Publisher statement

This work is made available according to the conditions of the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) licence. Full details of this licence are available at: https://creativecommons.org/licenses/by-nc-nd/4.0/

Publication date

2018-10-25

Notes

This paper is in closed access.

ISSN

1932-6254

eISSN

1932-7005

Language

en

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