Simple-Flow: A 3D-Printed Multiwell Flow Plate to Coculture Primary Human Lung Cells at the Air-Liquid Interface

Area of Science:

• Biomedical Engineering
• Cell Biology
• Pulmonology

Background:

• Traditional cell models (cell lines, animal models) for pulmonary disease research have limitations in recapitulating human lung physiology.
• Existing advanced in vitro models often face challenges with technical demands, low throughput, and reproducibility.
• There is a need for more accurate and practical in vitro models to improve the predictive outcomes of respiratory treatments.

Purpose of the Study:

• To introduce a novel, robust, and user-friendly fluidic device for advanced in vitro lung modeling.
• To demonstrate the capability of 3D-printing technology in creating customizable cell culture systems.
• To establish a more physiologically relevant in vitro model for studying pulmonary diseases.

Main Methods:

• Development of a biocompatible and customizable 3D-printed cell culture plate named Simple-Flow.
• Co-culture of human primary bronchial epithelial cells (hPBECs) at the air-liquid interface for mucociliary differentiation.
• Co-culture of human pulmonary microvascular endothelial cells (hMVECs) under flow conditions for up to 2 weeks.
• Utilizing 3D-printing for accessible and reproducible in vitro model fabrication.

Main Results:

• The Simple-Flow device demonstrated medium throughput, ease of manufacturing, and simplicity of setup.
• Successful co-culture of hPBECs and hMVECs, mimicking distinct lung compartments.
• Establishment of a functional air-liquid interface for hPBECs and continuous flow for hMVECs, enhancing model fidelity.
• Proof of principle for the versatility of 3D-printed devices in creating advanced in vitro lung models.

Conclusions:

• The Simple-Flow device represents a significant advancement in creating physiologically relevant in vitro lung models.
• 3D-printing technology offers a versatile platform for developing customizable and reproducible cell culture systems.
• This novel model holds promise for improving preclinical research in pediatric lung diseases and other pulmonary disorders.
• The developed model can enhance the predictive accuracy of respiratory treatment outcomes.