Scalable and ultrafast CAR-T cell production using microfluidics

Area of Science:

• Biotechnology
• Cell Therapy
• Cancer Research

Background:

• Chimeric antigen receptor T cell (CAR-T) therapy is a promising cancer treatment, especially for blood cancers.
• Current CAR-T manufacturing is slow (up to two weeks), leading to loss of vital naive-like T-cells (Tnlp) and reduced efficacy.
• This time-consuming process is a major bottleneck for widespread CAR-T therapy adoption.

Purpose of the Study:

• To develop and optimize a microfluidic device (MFD) for rapid CAR-T cell production.
• To integrate T-cell activation and lentiviral transduction into a single, ultrafast step (within 24 hours).
• To evaluate the MFD's efficiency in preserving T-cell phenotypes, particularly Tnlp.

Main Methods:

• Development and optimization of a novel microfluidic device (MFD) for CAR-T cell manufacturing.
• Integration of T-cell activation and lentiviral transduction within the MFD.
• Comparison of transduction rates and Tnlp percentages between the MFD and standard plate-based methods.

Main Results:

• The MFD achieved a 27% transduction rate in 24 hours, outperforming standard methods (17% and 8%).
• The ultrafast MFD protocol yielded approximately six times more CD3+ Tnlp compared to the standard 9-day protocol (18.07% vs. 3.97%).
• Significant preservation of CD4+ and CD8+ Tnlp was also observed in the MFD-produced CAR-T cells.

Conclusions:

• Microfluidic devices offer a scalable platform for streamlined CAR-T manufacturing.
• The developed MFD significantly reduces production time while preserving essential T-cell phenotypes.
• This technology has the potential to improve CAR-T therapy accessibility and clinical outcomes.