3D alveolar in vitro model based on epithelialized biomimetically curved culture membranes

Biomaterials, vol. 266, pp. 120436

Abstract

There is increasing evidence that surface curvature at a near-cell-scale influences cell behaviour. Epithelial or endothelial cells lining small acinar or tubular body lumens, as those of the alveoli or blood vessels, experience such highly curved surfaces. In contrast, the most commonly used culture substrates for in vitro modelling of these human tissue barriers, ion track-etched membranes, offer only flat surfaces. Here, we propose a more realistic culture environment for alveolar cells based on biomimetically curved track-etched membranes, preserving the mainly spherical geometry of the cells' native microenvironment. The curved membranes were created by a combination of three-dimensional (3D) micro film (thermo)forming and ion track technology. We could successfully demonstrate the formation, the growth and a first characterization of confluent layers of lung epithelial cell lines and primary alveolar epithelial cells on membranes shaped into an array of hemispherical microwells. Besides their application in submerged culture, we could also demonstrate the compatibility of the bioinspired membranes for air-exposed culture. We observed a distinct cellular response to membrane curvature. Cells (or cell layers) on the curved membranes reveal significant differences compared to cells on flat membranes concerning membrane epithelialization, areal cell density of the formed epithelial layers, their cross-sectional morphology, and proliferation and apoptosis rates, and the same tight barrier function as on the flat membranes. The presented 3D membrane technology might pave the way for more predictive barrier in vitro models in future.

Authors 13

1. Danielle Baptista

   Maastricht University

   Affiliation as printed

   MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands

2. Liliana Moreira Teixeira

   Maastricht University · University of Twente

   Affiliation as printed

   MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands; Department of Developmental BioEngineering, Technical Medical Centre, University of Twente, Drienerlolaan 5, 7522 NB, Enschede, the Netherlands

   Department of Developmental BioEngineering, Technical Medical Centre, University of Twente, Drienerlolaan 5, 7522 NB, Enschede, the Netherlands

   MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands

3. Zeinab Niloofar Tahmasebi Birgani

   Maastricht University

   Affiliation as printed

   MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands

4. Sander van Riet Aachen Dept of Pulmonology, Leiden University Medical Center

   Leiden University Medical Center

   Affiliation as printed

   Department of Pulmonology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA, Leiden, the Netherlands

5. Thijs Pasman

   University of Twente

   Affiliation as printed

   Department of Biomaterials Science and Technology, Technical Medical Centre, University of Twente, Drienerlolaan 5, 7522 NB, Enschede, the Netherlands

6. André A. Poot

   University of Twente

   Affiliation as printed

   Department of Biomaterials Science and Technology, Technical Medical Centre, University of Twente, Drienerlolaan 5, 7522 NB, Enschede, the Netherlands

7. Dimitrios F. Stamatialis

   University of Twente

   Affiliation as printed

   Department of Biomaterials Science and Technology, Technical Medical Centre, University of Twente, Drienerlolaan 5, 7522 NB, Enschede, the Netherlands

8. Robbert J. Rottier

   Erasmus MC · Erasmus MC - Sophia Children’s Hospital · Erasmus University Rotterdam

   Affiliation as printed

   Department of Pediatric Surgery/Cell Biology, Erasmus (University) Medical Center - Sophia Children's Hospital, Doctor Molewaterplein 40, 3015 GD, Rotterdam, the Netherlands

   Department of Pediatric Surgery/Cell Biology, Erasmus (University) Medical Center – Sophia Children's Hospital, Doctor Molewaterplein 40, 3015 GD, Rotterdam, the Netherlands

9. Pieter Sicco Hiemstra Aachen Dept of Pulmonology, Leiden University Medical Center

   Leiden University Medical Center

   Affiliation as printed

   Department of Pulmonology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA, Leiden, the Netherlands

10. Pamela Habibović

    Maastricht University

    Affiliation as printed

    MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands

11. Clemens van Blitterswijk

    Maastricht University

    Affiliation as printed

    MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands

12. Stefan Giselbrecht

    Maastricht University

    Affiliation as printed

    MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands

13. Roman K. Truckenmüller corresponding

    Maastricht University

    Affiliation as printed

    MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands. Electronic address: r.truckenmuller@maastrichtuniversity.nl

    MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Universiteitssingel 40, 6229 ER, Maastricht, the Netherlands