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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier Applied Science
2017
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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. |
format | Online Article Text |
id | pubmed-5615110 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Elsevier Applied Science |
record_format | MEDLINE/PubMed |
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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