From previous work, we have shown the efficacy of hydroporation in the generation of functional CAR+ T-cells, with higher yields and similar functionality to those generated by conventional electroporation. Here, we show that hydroporation can be used to generate universal CAR+T cells through multiplex editing, which are as functional as those generated by nucleofection but with greater viability and cell number. 

Recent studies suggest a strong connection between CAR-T proliferation, how long they last inside the patient’s body, and how well the patient responds to the treatment [3,16]. To better understand the connection, scientists have examined the ability of CAR-T cells to activate the appropriate immune response while simultaneously avoiding activation-induced cell death (AICD) [17,18].

The team used CAR-T cells modified with hydroporation and nucleofection by varying the number of CAR-T cells matched with a single cancer cell (effector-to-target ratio) to see if one method performed better. They found that hydroporated CAR-T cells were superior in all tested E:T ratios.

Even at a ratio of 1 CAR-T cell to 8 cancer cells, the hydroporated CAR-T cells were able to eliminate more than 90% of the cancer cells, whereas the nucleofected CAR-T cells (Nu-CAR-T) only managed to clear 52% of the cancer cells at the same ratio.

Reliable and effective T cell culture is critical to developing and testing novel CAR-T therapies. It is also a monotonous, error prone, and time consuming process when performed manually. Monomer Bio and Indee Labs have built a fully-automated work cell to address these problems. Herein, we compare the expansion of human primary T cells cultured by hand versus by the Monomer work cell.

Cyclic peptides can target specific interactions between proteins inside of cells, something that’s quite challenging with drugs under normal circumstances. Unfortunately, these peptides typically can’t enter cells on their own. Without intervention, studying their functionality is difficult at best. Here is how one team screened hundreds of different cyclic peptide conditions with Hydropore in minutes.

Hydropore™ is a novel, non-viral delivery platform now available for Research Use Only (RUO). Hydropore™ offers researchers a reliable alternative to electroporartion, and can rapidly and efficiently introduce nucleic acids, proteins and gene-editing complexes into cells for a variety of applications.

The most problematic step in GMCT manufacturing is the intracellular delivery of nucleic acids via transfection or transduction for expression of the CAR by the T cell. Current methods require significant hands-on time during production and require extensive intra- and post-production safety testing to avoid infusion of replication competent viruses at the time of therapy administration.

Microfluidic vortex shedding (µVS) has been shown to enable the efficient delivery of constructs like mRNA, plasmid DNA and Cas9 RNPs (CRISPR) to primary human T cells. But, what is µVS? And, how does µVS enable intracellular delivery of constructs to primary cells like activated T cells?

In this blog post, we will explore the advantages and limitations of some of the most common classes of therapeutics:  small molecules, macromolecules, nanoparticles, and chimeric antigen receptor T-cells.