Introducing Hydropore for Cell Biology & Cell Therapy Research
Developing a scalable and efficient transfection method is crucial in accelerating the discovery, development, and manufacturing of gene-modified cell therapies. Researchers need a reliable alternative that can rapidly and efficiently introduce nucleic acids, proteins and gene-editing complexes into cells for a variety of applications. However, current transfection methods such as electroporation have seen less success with transfection due to high toxicity and other limitations.
Indee Lab’s non-viral delivery platform, Hydropore™, is a robust solution that bypasses the limitations and costs of electroporation, viral transduction, and other non-viral transfection methods. Hydropore™ is being developed in collaboration with 3 of the top 10 pharmaceutical companies, various biotechs and leading research institutions including UC San Francisco, Stanford, Medical University of South Carolina, the National Cancer Institute and the National Institute of Allergy and Infectious Disease among others.
In this post, we provide an overview of Hydropore™ and the unique advantages it can bring to your research over electroporation like improved cell quality and lower cost per experiment. We also discuss the current applications and technical limitations of Hydropore™ along with its simple technology transfer process.
What is Hydropore™?Hydropore™ is a novel, non-viral delivery platform currently available for Research Use Only (RUO). This technology uses hydrodynamic conditions termed microfluidic vortex shedding (µVS) that can be coupled to a brief electric field to gently permeabilize cells, allowing for the quick and efficient delivery of constructs like gene-editing complexes and nucleic acids into cells.
Spacing between posts, along with a brief electric field, allows for T cells to flow through without significant physical deformation or perturbation of the cell state. The vortices created by the posts apply hydrodynamic forces to the cell membrane which cause temporary pores allowing for the diffusion and electrophoretic transfer of constructs into cells. Cells rapidly recover, exit the device, and are collected for downstream applications. Typically, each device can process 1~100 million cells in 1~30 seconds.
Figure 1 Illustration of the µVS intracellular delivery mechanism
Hydropore™ is shown to have a similar yield to electroporation but with improved cell quality and no need for multiple cuvettes. This technology bypasses existing limitations of electroporation methods by using GMP-grade OptiMEM media and applying just a brief electric field, allowing for a gentler system with a larger processing window (>1 hour). As a result, cells demonstrate increased viability, improved proliferation and preserved cell function.
Hydropore™ has a simple installation process with a nearly identical workflow to that of electroporation. For experiments that require multiple electroporation cuvettes, Hydropore™ leads to a lower cost per experiment relative to popular RUO electroporation systems. Ultimately, researchers are able to produce higher quality cells at a competitive price (Tables 1 & 2).
Table 1 Comparison of Hydropore™ to an RUO electroporation system. Assumes 100 µl cuvette and 50 million to 100 million cells per mL.
Table 2 Cost comparison per experiment of Hydropore™ to an RUO electroporation system. Assumes 100 µl cuvette and 50 million cells per mL.
Hydropore™ is currently available for Research Use Only (RUO) and is verified or in development for immune cells including T cells, Tregs and TILs, PBMCs, Natural Killer (NK) cells, cell models such as HEK293Ts & Jurkats and more. This platform is also verified for the delivery of CRISPR-Cas9 and mRNA to donor immune cells, specifically in primary human activated CD3+ T cells.
For areas where we do not currently have existing datasets, Hydropore™ works well for actively dividing, suspended or trypsinized adherent cells that are 8~15 µm in diameter. Hydropore™ also delivers membrane impermeable small molecules and Cas9 RNPs where electroporation editing efficiencies are >20% and
Figure 2 Workflow comparison of Hydropore™ to an RUO electroporation system.
The entire workflow of Hydropore™ is nearly identical to that of electroporation. Those with aseptic technique and electroporation experience can be easily onboarded from the first experiment. Our team provides an overview of the entire processing protocol along with video demonstrations and tech transfer documents. Scientists simply need compressed nitrogen close enough to the hood and will be provided with the right regulator and verified reagents like eGFP mRNA and OptiMEM to get started.
After that, scientists are able to pursue their research interests within the technical limitations described in the previous section.
Figure 3 Hydropore™ transfection protocol
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