Development of Cell-IN technology for microplate and flow cytometry applications

How to Penetrate the Inside of a Cell?

Every cell in the human body is enclosed by a cell membrane. This membrane acts as a protective shield against the external environment, making its penetration a significant hurdle and a limiting factor in most medical and biological research. Aneta Magiera, M.Sc. Eng., from the Institute of Physical Chemistry of the Polish Academy of Sciences in Warsaw, together with her research team, has developed Cell-IN—a reagent that enables simple and rapid crossing of the cell membrane. This allows for the delivery of various desired compounds into the cell, ranging from polymers, proteins, and nanoparticles to nucleic acids. Through the Proof of Concept project, funded by the European Funds for a Modern Economy 2021–2027 (FENG) program and implemented by the Foundation for Polish Science (FNP), she aims to optimize the workflow for this reagent to ensure high-throughput processing of large volumes of analyses within short timeframes.

"Cell-IN is a polymeric reagent that, when dissolved in a medium tailored to specific target cells, creates a high osmotic pressure solution. The core mechanism for delivering a target compound into the cell using Cell-IN relies on the rupture of intracellular vesicles triggered by osmotic shock. Metaphorically, Cell-IN acts as an 'airplane' that provides fast and effortless transport for passengers (the molecules) from, for instance, Poland to the United States (the human cells), easily crossing the U.S. border (the cell membrane). Crucially, it is a versatile reagent capable of introducing a wide variety of compounds into cells," says Aneta Magiera, M.Sc. Eng.

Although Cell-IN is already commercially available, the experimental workflow requires optimization for studies examining large cell populations rather than single cells. These high-throughput experiments typically rely on standard laboratory equipment such as flow cytometers and multi-well microplate readers. Adapting Cell-IN technology for compatibility with these instruments will enhance its competitive edge over alternative intracellular delivery methods, such as microinjection or electroporation.

"The research planned within the Proof of Concept project will drive the development of Cell-IN technology and advance its path toward further commercialization by confirming its deployment potential. End-users of the optimized Cell-IN protocol will include individual academic researchers requiring cell membrane penetration using 96-well culture plates or flow cytometry techniques, as well as pharmaceutical and biotechnology companies and chemical reagent manufacturers," explains Aneta Magiera, M.Sc. Eng.

The Cell-IN solution offers broad potential applications, including the targeted and straightforward intracellular delivery of therapeutics, such as anticancer compounds. The team plans to investigate using Cell-IN for nucleic acid delivery to facilitate cellular modification for therapeutic purposes.

Aneta Magiera, M.Sc. Eng. (Karpińska), completed her Bachelor's and Master's degrees in Biotechnology (specializing in Drugs and Cosmetics) at Warsaw University of Technology. In her doctoral research, she investigates the intracellular uptake of anticancer drugs in living cells using Fluorescence Correlation Spectroscopy (FCS). In 2023, she received a START stipend from the Foundation for Polish Science, ranking among four laureates whose research achievements were recognized as outstanding by competition reviewers. She represented Poland at the global finale of Falling Walls Lab 2024 in Berlin and at the 73rd Lindau Nobel Laureate Meeting. She is also a alumna of leadership and entrepreneurship programs designed for early-career scientists, including the international Excellerator 2023 program hosted by the University of Cambridge and Global Biotech Revolution, as well as the Shesnnovation program organized by the Perspektywy Women in Tech Foundation. She is actively involved in several research projects in collaboration with external institutions, including the University of Utah in the USA.