On-Chip Neural Induction Boosts Neural Stem Cell Commitment: Toward a Pipeline for iPSC-Based Therapies

Area of Science:

• Biotechnology
• Stem Cell Biology
• Regenerative Medicine

Background:

• Clinical translation of induced pluripotent stem cells (iPSCs) is hampered by costly, time-consuming, and non-standardized generation workflows.
• Developing efficient and scalable methods for iPSC generation and differentiation is crucial for personalized therapeutics.

Purpose of the Study:

• To present a simplified, cost-effective microfluidic approach for reprogramming fibroblasts into iPSCs and differentiating them into neural stem cells (NSCs).
• To evaluate the efficiency and quality of iPSCs and NSCs generated using microfluidic technology compared to conventional methods.

Main Methods:

• Utilized microfluidic chips and microphysiological technology for fibroblast reprogramming into iPSCs.
• Performed subsequent differentiation of iPSCs into NSCs within the microfluidic system.
• Compared reprogramming and neural induction efficiency using bulk RNA sequencing and pathway enrichment analysis.

Main Results:

• Microfluidic reprogramming achieved a 100-fold reduction in reagents and a ninefold reduction in input cells.
• Generated iPSCs exhibited upregulated pluripotency markers and downregulated fibroblast markers, comparable to conventional methods.
• Differentiated NSCs showed upregulated neuroectodermal markers and enhanced neural stem cell lineage commitment in microfluidic chips.

Conclusions:

• The microfluidic approach offers a cost-effective and efficient pipeline for generating iPSCs and NSCs.
• This method is compliant with current good manufacturing practices, supporting clinical translation for personalized therapeutics.
• Microfluidic confinement enhances neural stem cell development and commitment, offering advantages for nervous system development applications.