Establishing a Three-Dimensional Coculture Module of Epithelial Cells Using Nanofibrous Membranes

Area of Science:

• Biomaterials Science
• Cell Biology
• Tissue Engineering

Background:

• Epithelial cell culture faces challenges like dedifferentiation and loss of function.
• Biomimetic three-dimensional (3D) cell culture offers enhanced efficiency and physiological relevance.
• Advanced culture systems are needed to maintain epithelial cell integrity and function.

Purpose of the Study:

• To introduce and validate a two-layered co-culture system for cultivating epithelial cells and fibroblasts.
• To utilize polyvinyl alcohol (PVA) and poly(ε-caprolactone) (PCL) nanofibrous membranes as physiologically relevant scaffolds.
• To investigate paracrine signaling and cellular behavior in a biomimetic 3D environment.

Main Methods:

• Fabrication of PVA and PCL nanofibrous membranes (NMs) via electrospinning.
• Co-culturing lung epithelial cells on PVA NM and fibroblasts on PCL NM in an insert system.
• Analysis using confocal microscopy (with Z-stacking), scanning electron microscopy (SEM), and Cell Counting Kit-8 (CCK-8) assay.

Main Results:

• The system successfully cultured epithelial cells as tissue-like layers and fibroblasts in a 3D environment.
• Confocal microscopy and SEM provided insights into cell distribution, tight junction integrity, and morphology.
• CCK-8 assays revealed cell growth rates and potential synergistic effects in co-culture.

Conclusions:

• The developed insert co-culture system effectively supports simultaneous culture of epithelial cells and fibroblasts.
• This biomimetic approach is valuable for studying paracrine signaling in contexts like tissue regeneration and drug testing.
• The system offers a robust platform for physiological and pharmacological research involving epithelial tissues.