Revolutionary Laser-Based Platform Boosts CAR T Cell Manufacturing and Performance

A team of researchers from the Keck School of Medicine at USC has introduced an innovative analytical platform that significantly advances the study and production of chimeric antigen receptor (CAR) T cells. This cutting-edge system employs spectral flow cytometry, a laser-based technology capable of analyzing multiple cellular features simultaneously, providing a comprehensive view of CAR T cell characteristics and their evolution during manufacturing.

Using this platform, scientists have uncovered that CAR T cells are most potent when cultured for five days, as opposed to the traditional ten-day expansion. Early-stage cells exhibited more stem-like properties and higher metabolic activity, traits associated with improved long-term cancer-fighting efficacy. This insight allows for more precise timing in cell manufacturing to enhance therapy outcomes.

The study, published in the 25th-anniversary issue of 'Molecular Therapy', emphasizes the importance of understanding cellular fingerprints—the unique expression of surface markers—to refine and optimize CAR T cell therapies. The novel platform can analyze 36 markers on single cells at once, offering a holistic view that surpasses conventional methods limited to fewer markers.

The system begins by tagging cells with fluorescent markers that bind to specific molecules, then passing them through a spectral flow cytometer. Lasers excite these markers, emitting light that is detected and measured, revealing detailed information about cell activation, metabolism, memory, and cytotoxic ability. Researchers carefully selected the markers for their relevance to anti-cancer activity and ensured accurate detection through sophisticated mathematical models.

In their experiments, the team compared CAR T cells on days five and ten of expansion, finding that early cells resemble stem-like, highly metabolic cells better suited for durable cancer eradication. This discovery could inform manufacturing protocols, making the production process more tailored and effective.

Beyond manufacturing, the platform holds potential for a range of applications, including comparing gene editing techniques, analyzing different engineering platforms, and identifying biomarkers predictive of patient responses. Clinical trial centers could utilize this technology to monitor CAR T cell evolution in real time, providing critical insights into long-term success and treatment optimization.

"This technology is just beginning to unlock the potential of cellular therapies," said senior author Mohamed Abou-el-Enein. "Our scalable, collaborative platform paves the way for next-generation immunotherapies, accelerating development and improving patient outcomes."

The research team comprises experts from USC and CHLA, including Amaia Cadinanos-Garai, Christian L. Flugel, Anson Cheung, Enzi Jiang, and Alix Vassie. Their work opens new avenues for understanding and enhancing CAR T cell therapies, promising improved treatments for cancers resistant to conventional methods.

For more information, see the full study in 'Molecular Therapy'.

source: https://medicalxpress.com/news/2025-05-laser-based-analytical-platform-car.html