NGN2-based neuronal programming of hiPSCs in an automated microfluidic platform

Area of Science:

• Stem Cell Biology
• Neuroscience
• Bioengineering

Background:

• Induced pluripotent stem cells (iPSCs) offer a patient-specific in vitro cell source for disease modeling.
• Lab-on-a-chip technology provides advanced in vitro models with controlled microenvironments.
• Automated microfluidic platforms enhance throughput and standardization for biological assays.

Purpose of the Study:

• To develop an automated, user-friendly microfluidic platform for efficient human iPSC-to-neuron conversion.
• To address challenges in robustness and ease of use for lab-on-a-chip systems in biological research.
• To create a reliable in vitro model for neurological disease research and drug discovery.

Main Methods:

• Utilized multilayer soft-lithography for platform fabrication and assembly.
• Implemented automated operations including cell seeding, medium exchange, and doxycycline-induced neuronal differentiation.
• Integrated immunofluorescence assays for analysis of neuronal marker expression and calcium signaling.

Main Results:

• Achieved high-throughput, efficient, and homogenous conversion of hiPSCs to neurons within 10 days.
• Demonstrated successful neuronal differentiation via Neurogenin 2 (NGN2) overexpression.
• Confirmed neuronal identity through MAP2 expression and functional calcium signaling.

Conclusions:

• The automated neurons-on-chip platform facilitates rapid and standardized neuronal differentiation from hiPSCs.
• This system overcomes key limitations in current lab-on-a-chip technologies for biological applications.
• The developed model offers a robust tool for in vitro neurological disease modeling and preclinical drug screening.