Non-Invasive Quality Control of Organoid Cultures Using Mesofluidic CSTR Bioreactors and High-Content Imaging.

Area of Science:

• Neuroscience
• Biotechnology
• Bioengineering

Background:

• Human brain organoids model in vivo brain function and neurological diseases.
• Current 3D organoid cultures face challenges with long growth times and sample heterogeneity, limiting their application.
• Robust and reproducible methods are needed for long-term organoid culturing and quality control.

Purpose of the Study:

• To develop an integrated platform for robust, long-term culturing of 3D brain organoids.
• To improve nutrient delivery and enable non-invasive quality control of organoids.
• To establish reproducible culture standards for 3D cellular systems.

Main Methods:

• Designed a mesofluidic bioreactor based on reaction-diffusion scaling theory for efficient media exchange.
• Integrated the bioreactor with longitudinal tracking and machine learning for non-invasive organoid quality control.
• Performed transcriptome analyses to assess organoid development and cell viability.

Main Results:

• The mesofluidic bioreactor facilitated robust media exchange for long-term organoid culture.
• The integrated platform enabled non-invasive quality control and sample pre-selection.
• Transcriptome data indicated enhanced organoid development and reduced cell death in the bioreactor.

Conclusions:

• The developed integrated platform enables robust and reproducible long-term culturing of 3D brain organoids.
• This platform provides a generalizable tool for establishing consistent standards in 3D cellular systems.
• The system has broad applications in neurological disease modeling and therapeutic screening.