Microfluidic platforms for CAR-T cell engineering: a multidisciplinary review on potential next generation intracellular delivery method
10 affiliations
- 1Chemistry and Molecular Physics Department, University of São Paulo, São Carlos, Brazil
- 2Brazilian Nanotechnology National Laboratory, Brazilian Center for Research in Energy and Materials, Campinas, Brazil
- 3Micromanufacturing Laboratory, Institute for Technological Research, São Paulo, Brazil
- 4Microelectronics Department, University of São Paulo, São Paulo, Brazil
- 5Basque Center for Applied Mathematics (BCAM), Bilbao, Spain
- 6Institute of Technology in Immunobiologicals (Bio-Manguinhos), Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, Brazil
- 7Cell and Gene Therapy Program - Research Coordination, Brazilian National Cancer Institute (INCA), Rio de Janeiro, Brazil
- 8Vice-Presidency of Research and Biological Collections (VPPCB), Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, Brazil
- 9Pasteur Fiocruz Center on Immunology and Immunotherapy, Ceará, Brazil
- 10Nanotechnology Engineering Department (PENt), Alberto Luiz Coimbra Institute for Graduate Studies and Research in Engineering (COPPE), Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil
Abstract
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.
Graphical Abstract:
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BibTeX
@article{andreotti2026microfluidic,
title = {{Microfluidic platforms for CAR-T cell engineering: a multidisciplinary review on potential next generation intracellular delivery method}},
author = {Isabela Aparecida de Araujo Andreotti and Gabriel Griep and Edgar Andrés Patiño Nariño and Leonardo Ribeiro Batista-Silva and Mario Ricardo Góngora-Rubio and Martín Hernán Bonamino and Tiago Albertini Balbino},
journal = {Journal of Translational Medicine},
year = {2026},
month = jul,
doi = {10.1186/s12967-026-08388-4},
url = {https://doi.org/10.1186/s12967-026-08388-4},
note = {CC BY-NC-ND 4.0},
}
RIS
TY - JOUR
TI - Microfluidic platforms for CAR-T cell engineering: a multidisciplinary review on potential next generation intracellular delivery method
AU - Isabela Aparecida de Araujo Andreotti
AU - Gabriel Griep
AU - Edgar Andrés Patiño Nariño
AU - Leonardo Ribeiro Batista-Silva
AU - Mario Ricardo Góngora-Rubio
AU - Martín Hernán Bonamino
AU - Tiago Albertini Balbino
PY - 2026
DA - 2026/07/29
JO - Journal of Translational Medicine
DO - 10.1186/s12967-026-08388-4
UR - https://doi.org/10.1186/s12967-026-08388-4
AB - 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. Graphical Abstract:
SN - 1479-5876
ER -