Aggregates-on-chip: a microfluidic device for crowding plant cells in culture

Area of Science:

• Plant Cell Biology
• Microfluidics
• Biotechnology

Background:

• Microfluidic devices offer controlled environments for studying cellular properties.
• Understanding cell-cell contact and microenvironmental influences is crucial for plant cell growth.
• Existing microfluidic systems often lack the ability to facilitate cell-cell contact for long-term plant cell studies.

Purpose of the Study:

• To develop a novel microfluidic device enabling sustained cell-cell contact and long-term imaging of plant protoplasts.
• To create a constrained microenvironment that mimics natural plant tissue interactions.
• To investigate the impact of cell-cell contact and microenvironmental cues on plant protoplast behavior.

Main Methods:

• Development of a microfluidic device utilizing arrays of cylindrical cups to form stable protoplast aggregates.
• Incorporation of a porous micro-post design to ensure sustained media flux and prevent hypoxic conditions.
• Implementation of sterile culture conditions with convenient media switching for various assays.

Main Results:

• The microfluidic platform successfully maintained protoplast viability for over two weeks in Arabidopsis thaliana and Zea mays.
• Observed cell growth, cell wall formation, and cell divisions within the confined aggregates.
• Demonstrated significantly higher cell division rates in crowded aggregates compared to isolated cells.

Conclusions:

• The developed aggregates-on-chip platform is a valuable tool for long-term, live-imaging studies of plant protoplast behavior.
• The device facilitates the investigation of how physical confinement, perfusion, and chemical signals impact plant cell growth, regeneration, division, and stress responses.
• This technology advances the study of cell-cell interactions and microenvironmental effects in plant biology.