Jason N. Belling, Joshua A. Jackman, Liv K. Heidenreich, Lisa M. Kawakami, Thomas Young, Leonardo Scarabelli, Chuanzhen Zhao, Nam-Joon Cho, Steven J. Jonas, and Paul S. Weiss
Gene-editing tools are leading to a new era of personalized medicine, including gene therapy for genetic blood disorders and CAR-T cell therapies for blood cancers. Because of the limitations of lentivirus-based gene delivery, there is strong demand for the development of scalable technologies to edit the genomes of individual cells with high efficiency and speed. Towards this goal, constrictive channel microfluidic systems have emerged as a promising technology that facilitates intracellular delivery of biomolecular cargo using rapid cell squeezing to achieve membrane disruption. As microfluidic technologies transition from conceptual prototypes to functional tools, there is a need to develop next-generation platforms that are able to process cells in great number while avoiding clogging. We report a new method to deliver biomolecules intracellularly using PDMS constrictive channels that leads to dramatically less cell clogging and device failure. Further, we report a versatile method to passivate microfluidic channels based on noncovalent lipid bicelle technology, leading to dramatic improvements in blocking efficiency against nonspecific protein adsorption and cell adhesion as compared to covalent polymer options.