3D Printed Transwell Microfluidic Devices for Epithelial Cell Culture with Shear Stress.

Area of Science:

• Biotechnology
• Biomaterials Engineering
• Cell Biology

Background:

• Traditional cell culture models often fail to replicate the complex microenvironment and physiological conditions of in vivo systems.
• Developing advanced cell culture platforms is crucial for accurate drug discovery and disease modeling.

Purpose of the Study:

• To develop and validate a 3D-printed microfluidic transwell system for robust, long-term cell culture under recirculating flow.
• To create a more physiologically relevant model for studying epithelial barrier function and drug transport.

Main Methods:

• Fabrication of a microfluidic device using 3D printing, electrospun collagen scaffolds, and laser-cut Teflon membranes.
• Culture of Madin-Darby canine kidney (MDCK) cells to form an epithelial monolayer within the device.
• Characterization of cell morphology, barrier function (TEER), gene expression, and drug permeability using various analytical techniques.

Main Results:

• Successful fabrication of a microfluidic transwell system with stable fluidic connections and a printed reservoir for cell seeding.
• Demonstration of MDCK cell monolayer integrity and response to continuous flow, enabling monitoring of transport.
• Accurate assessment of drug permeability for caffeine and digoxin, yielding results comparable to in vivo studies.

Conclusions:

• The 3D-printed microfluidic transwell system provides a robust and in vivo-like platform for long-term cell culture and transport studies.
• This technology holds significant potential for advancing drug development and understanding biological barrier functions.
• The system's ability to mimic physiological conditions makes it a valuable tool for preclinical research.