High-throughput continuous-flow microfluidic electroporation of mRNA into primary human T cells for applications in cellular therapy manufacturing.
Sci Rep
; 10(1): 18045, 2020 10 22.
Article
em En
| MEDLINE
| ID: mdl-33093518
ABSTRACT
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[Formula see text]-10[Formula see text] cells, and that it is well-suited to scale for production of trillions of cells to support emerging allogeneic therapies.
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
RNA Mensageiro
/
Linfócitos T
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Transfecção
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Técnicas de Transferência de Genes
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Eletroporação
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Microfluídica
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Terapia Baseada em Transplante de Células e Tecidos
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Sistemas CRISPR-Cas
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Edição de Genes
Limite:
Humans
Idioma:
En
Revista:
Sci Rep
Ano de publicação:
2020
Tipo de documento:
Article
País de afiliação:
Estados Unidos