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

Baptista, Danielle; Teixeira, Liliana Moreira; Birgani, Zeinab Tahmasebi; Riet, Sander van; Pasman, Thijs; Poot, Andreas A.; Stamatialis, Dimitrios; Rottier, Robbert J.; Hiemstra, Pieter S.; Habibović, Pamela; Blitterswijk, Clemens van; Giselbrecht, Stefan; Truckenmüller, Roman

doi:10.1016/j.biomaterials.2020.120436

Cited by 39 publications

(46 citation statements)

References 67 publications

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“…As shown in this study, rendering the cell culture surfaces with ECM-based proteins had a large impact on cells’ responses. Physiologically relevant cell culture platforms have been developed and showed that surface curvature influences alveolar cell behavior [ 9 , 54 ]. In these studies, rigid synthetic polymers have been used to mimic the desired curvature.…”

Section: Discussionmentioning

confidence: 99%

A Robust Protocol for Decellularized Human Lung Bioink Generation Amenable to 2D and 3D Lung Cell Culture

Dabaghi

Saraei

Carpio

et al. 2021

Cells

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Decellularization efforts must balance the preservation of the extracellular matrix (ECM) components while eliminating the nucleic acid and cellular components. Following effective removal of nucleic acid and cell components, decellularized ECM (dECM) can be solubilized in an acidic environment with the assistance of various enzymes to develop biological scaffolds in different forms, such as sheets, tubular constructs, or three-dimensional (3D) hydrogels. Each organ or tissue that undergoes decellularization requires a distinct and optimized protocol to ensure that nucleic acids are removed, and the ECM components are preserved. The objective of this study was to optimize the decellularization process for dECM isolation from human lung tissues for downstream 2D and 3D cell culture systems. Following protocol optimization and dECM isolation, we performed experiments with a wide range of dECM concentrations to form human lung dECM hydrogels that were physically stable and biologically responsive. The dECM based-hydrogels supported the growth and proliferation of primary human lung fibroblast cells in 3D cultures. The dECM is also amenable to the coating of polyester membranes in Transwell™ Inserts to improve the cell adhesion, proliferation, and barrier function of primary human bronchial epithelial cells in 2D. In conclusion, we present a robust protocol for human lung decellularization, generation of dECM substrate material, and creation of hydrogels that support primary lung cell viability in 2D and 3D culture systems

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Section: Discussionmentioning

confidence: 99%

A Robust Protocol for Decellularized Human Lung Bioink Generation Amenable to 2D and 3D Lung Cell Culture

Dabaghi

Saraei

Carpio

et al. 2021

Cells

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“…Developments in delivering more advanced membrane technologies are now seeing a significant push [77,125,129,132]. For example, combining a micro film (thermo)forming and ion track technology Baptista et al have introduced curved track-etched membranes, exemplifying spherical geometries reminiscent of acinar structures [133]. Typically, these and other "soft" membranes can be integrated into relatively large arrays supporting the prospect of large throughput assays at an ALI [86], but are however restricted to geometrical constructs that largely forego the intrinsic lung anatomy (i.e.…”

Section: Mechanical Strains Of the Acinar Lumenmentioning

confidence: 99%

Advanced human-relevant in vitro pulmonary platforms for respiratory therapeutics

Artzy‐Schnirman

Raviv

Flikshtain

et al. 2021

Advanced Drug Delivery Reviews

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Over the past years, advanced in vitro pulmonary platforms have witnessed exciting developments that are pushing beyond traditional preclinical cell culture methods. Here, we discuss ongoing efforts in bridging the gap between in vivo and in vitro interfaces and identify some of the bioengineering challenges that lie ahead in delivering new generations of human-relevant in vitro pulmonary platforms. Notably, in vitro strategies using foremost lung-on-chips and biocompatible "soft" membranes have focused on platforms that emphasize phenotypical endpoints recapitulating key physiological and cellular functions. We review some of the most recent in vitro studies underlining seminal therapeutic screens and translational applications and open our discussion to promising avenues of pulmonary therapeutic exploration focusing on liposomes. Undeniably, there still remains a recognized trade-off between the physiological and biological complexity of these novel in vitro lung models and their ability to deliver assays with throughput capabilities. The upcoming years are thus anticipated to see further developments in broadening the applicability of such in vitro systems and accelerating therapeutic exploration for drug discovery and translational medicine in treating respiratory disorders.

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“…main carina in Figure 1f-h). Our development of more realistic 3D in vitro airway morphologies is supported by recent studies where Calu-3 cells (as well as other epithelial cells) cultured on curved membranes and inside lumen exhibit distinct characteristics from traditional 2D monolayers, including cell density and shape, apoptotic ratios, and cross-sectional morphology 41,42 . Here, we specifically designed a planar bifurcating geometry to facilitate microscopy imaging by limiting the model's vertical dimension, whose 3D curvature of the inner lumen extends outside the focal depth under higher magnifications (Figure 1h).…”

Section: Development Of 3d Bronchial Epithelial Airway Modelmentioning

confidence: 64%

“…For example, lung-on-chips 21,59 have established an attractive strategy to study amongst other human lung inflammation and drug responses in vitro by recreating a human-relevant alveolar microenvironment. To date, some studies have investigated ventilator-induced lung injury (VILI) and modeled the downstream barotrauma effects of pressure and membranal stretching at the acinar microscales only 41,54 whereas convective flow phenomena occurring upstream at the macroscale (i.e. conducting airways) have been largely overlooked 60 .…”

Section: Discussionmentioning

confidence: 99%

“…We chose Calu-3, a well-established lung cell line for preclinical pulmonary research that grows confluent layers in immersed conditions and forms barrier junctions. Previous lung-on-chip models have recapitulated other essential aspects of epithelial cell function such as ciliation 61 and mucus 41 secretion by culturing primary cells at an air-liquid-interface, but have excluded physiologically-relevant flow conditions critical to our study. Mucus is an aqueous secretion that offers a critical protective lining to epithelial cells covering the in vivo airways and serves to clear contaminants upwards via ciliary motion.…”

Section: Discussionmentioning

confidence: 99%

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Ventilation-induced epithelial injury drives biological onset of lung traumain vitroand is mitigated with anti-inflammatory therapeutics

Nof

Artzy‐Schnirman

Bhardwaj

et al. 2021

Preprint

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Mortality rates among patients suffering from acute respiratory failure remain perplexingly high despite maintenance of blood homeostasis. The biotrauma hypothesis advances that mechanical forces from invasive ventilation trigger immunological factors that spread systemically. Yet, how these forces elicit an immune response remains unclear. Here we show that flow-induced stresses under mechanical ventilation can injure the bronchial epithelium of ventilated in vitro upper airway models and directly modulate inflammatory cytokine secretion associated with pulmonary injury. We identify site-specific susceptibility to epithelial erosion in airways from jet-flow impaction and measure an increase in cell apoptosis and modulated secretions of cytokines IL-6, 8 and 10. We find that prophylactic pharmacological treatment with anti-inflammatory therapeutics reduces apoptosis and pro-inflammatory signaling during ventilation. Our 3D in vitro airway platform points to a previously overlooked origin of lung injury and showcases translational opportunities in preclinical pulmonary research towards protective therapies and improved protocols for patient care.

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

Cited by 39 publications

References 67 publications

A Robust Protocol for Decellularized Human Lung Bioink Generation Amenable to 2D and 3D Lung Cell Culture

A Robust Protocol for Decellularized Human Lung Bioink Generation Amenable to 2D and 3D Lung Cell Culture

Advanced human-relevant in vitro pulmonary platforms for respiratory therapeutics

Ventilation-induced epithelial injury drives biological onset of lung traumain vitroand is mitigated with anti-inflammatory therapeutics

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