Protocol to generate a microfluidic vessels-on-chip platform using human pluripotent stem cell-derived endothelial...

Area of Science:

• Biotechnology and Biomedical Engineering
• Stem Cell Biology
• Vascular Biology

Background:

• Microfluidic platforms offer advanced models for studying cellular functions.
• Human pluripotent stem cell-derived endothelial cells (SC-ECs) are crucial for vascular research.
• Developing robust protocols for in vitro vascular models is essential for drug discovery and disease modeling.

Purpose of the Study:

• To present a detailed protocol for fabricating a microfluidic vessel-on-chip.
• To establish a method for culturing and utilizing human SC-ECs within the microfluidic system.
• To enable multi-omics analysis of vascular structures formed on-chip.

Main Methods:

• Manufacturing of a 3D-printed microfluidic chip.
• Generation and culturing of human pluripotent stem cell-derived endothelial cells (SC-ECs).
• Hydrogel patterning for vessel formation and cultivation within an open microfluidic chip.
• Procedures for retrieving cells and media for downstream multi-omics analysis.

Main Results:

• Successful production of a microfluidic vessel-on-chip platform.
• Demonstration of barrier-forming vessel creation using SC-ECs.
• Establishment of a method for sample retrieval for multi-omics analysis.

Conclusions:

• The presented protocol provides a reproducible method for creating a functional microfluidic vascular model.
• This platform facilitates advanced studies on vascular biology and endothelial cell function.
• The integration of multi-omics analysis capabilities enhances the utility of the vessel-on-chip system.