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Transfection in perfused microfluidic cell culture devices: A case study

Automated microfluidic devices are a promising route towards a point-of-care autologous cell therapy. The initial steps of induced pluripotent stem cell (iPSC) derivation involve transfection and long term cell culture. Integration of these steps would help reduce the cost and footprint of micro-sca...

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Autores principales: Raimes, William, Rubi, Mathieu, Super, Alexandre, Marques, Marco P.C., Veraitch, Farlan, Szita, Nicolas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier Applied Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5615110/
https://www.ncbi.nlm.nih.gov/pubmed/28989299
http://dx.doi.org/10.1016/j.procbio.2016.09.006
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author Raimes, William
Rubi, Mathieu
Super, Alexandre
Marques, Marco P.C.
Veraitch, Farlan
Szita, Nicolas
author_facet Raimes, William
Rubi, Mathieu
Super, Alexandre
Marques, Marco P.C.
Veraitch, Farlan
Szita, Nicolas
author_sort Raimes, William
collection PubMed
description Automated microfluidic devices are a promising route towards a point-of-care autologous cell therapy. The initial steps of induced pluripotent stem cell (iPSC) derivation involve transfection and long term cell culture. Integration of these steps would help reduce the cost and footprint of micro-scale devices with applications in cell reprogramming or gene correction. Current examples of transfection integration focus on maximising efficiency rather than viable long-term culture. Here we look for whole process compatibility by integrating automated transfection with a perfused microfluidic device designed for homogeneous culture conditions. The injection process was characterised using fluorescein to establish a LabVIEW-based routine for user-defined automation. Proof-of-concept is demonstrated by chemically transfecting a GFP plasmid into mouse embryonic stem cells (mESCs). Cells transfected in the device showed an improvement in efficiency (34%, n = 3) compared with standard protocols (17.2%, n = 3). This represents a first step towards microfluidic processing systems for cell reprogramming or gene therapy.
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spelling pubmed-56151102017-10-05 Transfection in perfused microfluidic cell culture devices: A case study Raimes, William Rubi, Mathieu Super, Alexandre Marques, Marco P.C. Veraitch, Farlan Szita, Nicolas Process Biochem Article Automated microfluidic devices are a promising route towards a point-of-care autologous cell therapy. The initial steps of induced pluripotent stem cell (iPSC) derivation involve transfection and long term cell culture. Integration of these steps would help reduce the cost and footprint of micro-scale devices with applications in cell reprogramming or gene correction. Current examples of transfection integration focus on maximising efficiency rather than viable long-term culture. Here we look for whole process compatibility by integrating automated transfection with a perfused microfluidic device designed for homogeneous culture conditions. The injection process was characterised using fluorescein to establish a LabVIEW-based routine for user-defined automation. Proof-of-concept is demonstrated by chemically transfecting a GFP plasmid into mouse embryonic stem cells (mESCs). Cells transfected in the device showed an improvement in efficiency (34%, n = 3) compared with standard protocols (17.2%, n = 3). This represents a first step towards microfluidic processing systems for cell reprogramming or gene therapy. Elsevier Applied Science 2017-08 /pmc/articles/PMC5615110/ /pubmed/28989299 http://dx.doi.org/10.1016/j.procbio.2016.09.006 Text en © 2016 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Raimes, William
Rubi, Mathieu
Super, Alexandre
Marques, Marco P.C.
Veraitch, Farlan
Szita, Nicolas
Transfection in perfused microfluidic cell culture devices: A case study
title Transfection in perfused microfluidic cell culture devices: A case study
title_full Transfection in perfused microfluidic cell culture devices: A case study
title_fullStr Transfection in perfused microfluidic cell culture devices: A case study
title_full_unstemmed Transfection in perfused microfluidic cell culture devices: A case study
title_short Transfection in perfused microfluidic cell culture devices: A case study
title_sort transfection in perfused microfluidic cell culture devices: a case study
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5615110/
https://www.ncbi.nlm.nih.gov/pubmed/28989299
http://dx.doi.org/10.1016/j.procbio.2016.09.006
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