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Updated: Oct 28, 2026
Cell Squeezing as a Robust, Microfluidic Intracellular Delivery Platform
Published on: November 7, 2013
Microfluidic platforms for CAR-T cell engineering: a multidisciplinary review on potential next generation
Isabela Aparecida de Araujo Andreotti1,2,3, Gabriel Griep4, Edgar Andrés Patiño Nariño5
1Chemistry and Molecular Physics Department, University of São Paulo, São Carlos, Brazil.
Background:
Chimeric antigen receptor T (CAR-T) cell therapies have transformed the treatment landscape for hematological malignancies, yet their large-scale manufacturing still faces significant technical, economic, and regulatory constraints. Microfluidic-mediated mechanical poration (MMP) has emerged as a promising non-viral strategy for intracellular delivery in cell engineering, offering high-throughput operation, reduced reagent consumption, and the possibility of preserving cell viability compared with conventional physical delivery methods.
Main Body:
This review provides a structured assessment of the main MMP modalities, including constriction-based squeezing, volume exchange convective transfection (VECT), vortex shedding, spiral/viscoelastic hydroporation, and droplet mechanoporation, highlighting how device geometry, flow regime, and deformation mechanism influence delivery efficiency and cell integrity. Across the studies analyzed, reported delivery efficiencies frequently range from 60% to >95%, with cell viabilities commonly above 80-90% and processing capacities reaching from approximately 1 × 106 to 2.5 × 108 cells/min, depending on the device design, cargo, cell type, and operating conditions. Compared with viral vectors, MMP avoids complex vector production and may reduce biosafety and manufacturing burdens, while compared with electroporation, it may offer advantages in continuous-flow processing, cytotoxicity reduction, and cell recovery.
Conclusions:
Microfluidic-mediated mechanical poration shows strong potential as a next-generation intracellular delivery strategy for CAR-T cell manufacturing, combining efficiency, scalability, and reduced cytotoxicity. However, the current evidence remains uneven: many studies still rely on model cell lines or surrogate cargos, and direct validation in primary human T cells using CAR-encoding mRNA, DNA, or CRISPR-Cas9 components remains limited. Further optimization, regulatory alignment, and GMP-compatible integration are required to translate MMP from high-performing laboratory prototypes into reliable components of next-generation CAR-T cell manufacturing workflows.
