Lab-on-a-Chip Device for T Cell Isolation and CART Cell Therapy

Engineered cellular treatment employing viral vectors is currently considered a therapeutic strategy for treating both malignant and non-malignant leukaemias. However, this method comes with significant limitations. A primary concern about viral vectors is immunogenicity, as host immune systems identify viral elements as foreign substances, triggering a defensive response that can decrease vector delivery effectiveness and cause adverse side effects with high toxicity.

On the other hand, in the last two decades, due to the rapid development of micro and nanotechnology integration with chemistry and biology, a new branch has begun, known as microfluidics or Lab-on-a-Chip devices. The device has proven to be a powerful tool for cellular analysis in a complex micro-environment with its strengths of minimum sample consumption and contamination. These devices can potentially manipulate and detect biosamples, reagents, or biomolecules in a micro- environment and precisely perform cellular analysis because of their real-time operations, range of designing, easy fluid control, monitoring, and programmatic switching. The devices are useful for cell trapping, manipulation, isolation, separation, and lysis.

Despite advances in the field of microfluidic or Lab-on-a-Chip-based devices, achieving high transfection efficiency in the suspension cell population remains a challenge. With growing demand for highly efficient and targeted intracellular delivery, microfluidic or Lab-on-a-Chip based near-infrared laser-activated photoporation has emerged as a promising strategy that integrates precision and minimal invasiveness. Photoporation achieves precise biomolecule delivery control through spatial and temporal resolution by creating temporary cell membrane pores using light–matter interactions to disrupt the plasma membrane, making them inherently less toxic compared to chemical or viral methods. This leads to high delivery efficiency and minimal cellular damage. The technique enables targeted delivery of diverse biomolecules such as nanoparticles, macromolecules, and genetic materials to various cell types, including primary and stem cells that are notoriously difficult to transfect

Here we plan to develop a microfluidics or Lab-on-Chip-based, near-infrared pulse laser-activated photoporation platform that offers numerous advantages, such as T cell isolation, enabling suspension cells transfection, and the conduction of multiple experiments within the same device. These emphasize the development of a highly efficient, low-cost therapeutic tool. We plan to investigate the transfection of CAR gene into primary T cells. Anti-CD19 CAR-T cells are now widely used to treat refractory and relapsed B-cell leukaemia and lymphoma. Anti-BCMA CAR-T therapy is once again being used to treat multiple myeloma. However, the high-throughput generation of CAR-T cells remains an industrial challenge for personalized therapy. We are hopeful that successful CAR-T therapy using our pulse laser activated photoporation device can open up new era for clinical trials.