High-throughput continuous-flow microfluidic electroporation of mRNA into primary human T cells for applications in cellular therapy manufacturing

Autor: Daniel K. Freeman, Vienna L. Mott, Nerses J. Haroutunian, Jenna L. Balestrini, Vishal Tandon, Ernest S. Kim, Andrew Czarnecki, Charles A Lissandrello, Aaron G Stoddard, Michaela Welch, Peter Hsi, Jonathan R. Coppeta, Jordan P. Chesin, Jose A. Santos, Deborah A Flusberg
Rok vydání: 2020
Předmět:
Zdroj: Scientific Reports, Vol 10, Iss 1, Pp 1-16 (2020)
Scientific Reports
ISSN: 2045-2322
DOI: 10.1038/s41598-020-73755-0
Popis: Implementation of gene editing technologies such as CRISPR/Cas9 in the manufacture of novel cell-based therapeutics has the potential to enable highly-targeted, stable, and persistent genome modifications without the use of viral vectors. Electroporation has emerged as a preferred method for delivering gene-editing machinery to target cells, but a major challenge remaining is that most commercial electroporation machines are built for research and process development rather than for large-scale, automated cellular therapy manufacturing. Here we present a microfluidic continuous-flow electrotransfection device designed for precise, consistent, and high-throughput genetic modification of target cells in cellular therapy manufacturing applications. We optimized our device for delivery of mRNA into primary human T cells and demonstrated up to 95% transfection efficiency with minimum impact on cell viability and expansion potential. We additionally demonstrated processing of samples comprising up to 500 million T cells at a rate of 20 million cells/min. We anticipate that our device will help to streamline the production of autologous therapies requiring on the order of 10$$^8$$ 8 –10$$^9$$ 9 cells, and that it is well-suited to scale for production of trillions of cells to support emerging allogeneic therapies.
Databáze: OpenAIRE