Microfluidic reprogramming to pluripotency of human somatic cells.

Area of Science:

• Stem Cell Biology
• Regenerative Medicine
• Microfluidics

Background:

• Human induced pluripotent stem cells (hiPSCs) hold promise for regenerative medicine and precision medicine.
• Current reprogramming methods are inefficient, costly, and labor-intensive, limiting clinical translation.
• A need exists for streamlined, cost-effective hiPSC generation protocols.

Purpose of the Study:

• To develop a highly efficient, cost-effective, and rapid protocol for reprogramming human somatic cells into hiPSCs.
• To leverage microfluidic technology to optimize the reprogramming process.
• To generate clinical-grade hiPSCs under xeno-free conditions.

Main Methods:

• Downscaled an 8-day mRNA transfection protocol using microfluidics.
• Utilized daily transfections of mRNA encoding reprogramming factors and immune evasion proteins.
• Cultured cells under xeno-free defined conditions within a microfluidic chamber.

Main Results:

• Achieved high-efficiency hiPSC generation in 15 days.
• Obtained up to 160 ± 20 hiPSC colonies per 27 mm² microfluidic chamber.
• Required only ~20 µL of medium daily, with a ~100-fold reduction in raw material costs.
• Generated hiPSC colonies did not require further stabilization due to mRNA's short half-life.

Conclusions:

• Microfluidic-based reprogramming offers a highly efficient and scalable method for hiPSC generation.
• This protocol significantly reduces costs and workload, facilitating clinical translation.
• The system enables simultaneous reprogramming of hundreds of cells under defined conditions, advancing stem cell research and applications.