Introducing Hydropore™ for Cell Biology & Cell Therapy Research
By Indee Labs
Developing a scalable and efficient transfection method is crucial in accelerating gene-modified cell therapies' discovery, development, and manufacturing. However, current methods, such as electroporation, have seen less success due to high toxicity and other limitations.
Researchers need a reliable alternative that can rapidly and efficiently introduce nucleic acids, proteins, and gene-editing complexes into cells for various applications.
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 with 3 of the top 10 pharmaceutical companies, 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.
This post provides an overview of Hydropore™ and its unique advantages for your research, 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 and a brief electric field allows 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
Key Benefits of Hydropore™
Hydropore™ has 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.
This allows 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 electroporation. For experiments that require multiple electroporation cuvettes, Hydropore™ has a lower cost per experiment relative to popular RUO systems. Ultimately, researchers can produce higher-quality cells at a competitive price (Tables 1 and 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.
Current Applications for Hydropore™
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 delivering CRISPR-Cas9 and mRNA to donor immune cells, specifically in primary human-activated CD3+ T-cells.
For areas where we do not currently have 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 <3 kb nucleic acids like mRNA and DNA nanoplasmids.
Due to the gentle nature of Hydropore™, it is successful for all fragile and hard-to-transfect cell types tested to date.
Interested in using Hydropore™ in your lab or developing Hydropore™ for other applications?
Figure 2 Workflow comparison of Hydropore™ to an RUO electroporation system.
Easy & Verified Technology Transfer
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 processing protocol, 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. After that, scientists can pursue their research interests.
Figure 3 Hydropore™ transfection protocol