Protocols to co-culture human primary lung cells in the simple-flow device

Area of Science:

• Pulmonary Medicine
• Cell Biology
• Biomaterials Engineering

Background:

• Human lung models are crucial for research but often lack detailed culture methodologies.
• Increasing complexity in models like co-cultures and lung-on-chip devices necessitates standardized protocols.

Purpose of the Study:

• To provide detailed protocols for isolating, maintaining, and co-culturing primary human lung cells.
• To describe the 3D printing and setup of a simple-flow device for lung cell culture.
• To enable more reproducible and complex human lung model development.

Main Methods:

• Isolation, maintenance, and co-culturing of primary human epithelial, endothelial, and mesenchymal cells from lung resections.
• 3D printing of a simple-flow device.
• Setup of the device with co-cultures under fluidic conditions.

Main Results:

• Successful isolation and co-culturing of primary human lung cells.
• Demonstration of a 3D-printed simple-flow device suitable for lung cell culture.
• Establishment of fluidic conditions for epithelial and endothelial cell co-cultures.

Conclusions:

• This work provides a comprehensive guide for creating advanced human lung models.
• The described methods facilitate the development of more physiologically relevant in vitro lung systems.
• Standardized protocols are essential for advancing lung research using complex cell culture models.