A cost-effective and efficient reprogramming platform for large-scale production of integration-free human induced pluripotent stem cells in chemically defined culture

Factors limiting the adoption of iPSC technology include the cost of developing lines and the time period that it takes to characterize and bank them, particularly when integration free, feeder free, and Xeno-free components are used. In this manuscript we describe our optimization procedure that en...

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Autores principales: Beers, Jeanette, Linask, Kaari L., Chen, Jane A., Siniscalchi, Lauren I., Lin, Yongshun, Zheng, Wei, Rao, Mahendra, Chen, Guokai
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4464084/
https://www.ncbi.nlm.nih.gov/pubmed/26066579
http://dx.doi.org/10.1038/srep11319
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author Beers, Jeanette
Linask, Kaari L.
Chen, Jane A.
Siniscalchi, Lauren I.
Lin, Yongshun
Zheng, Wei
Rao, Mahendra
Chen, Guokai
author_facet Beers, Jeanette
Linask, Kaari L.
Chen, Jane A.
Siniscalchi, Lauren I.
Lin, Yongshun
Zheng, Wei
Rao, Mahendra
Chen, Guokai
author_sort Beers, Jeanette
collection PubMed
description Factors limiting the adoption of iPSC technology include the cost of developing lines and the time period that it takes to characterize and bank them, particularly when integration free, feeder free, and Xeno-free components are used. In this manuscript we describe our optimization procedure that enables a single technician to make 20–40 lines at a time in a 24–96 well format in a reliable and reproducible fashion. Improvements spanned the entire workflow and included using RNA virus, reducing cytotoxicity of reagents, developing improved transfection and freezing efficiencies, modifying the manual colony picking steps, enhancing passaging efficiency and developing early criteria of success. These modifications allowed us to make more than two hundred well-characterized lines per year.
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spelling pubmed-44640842015-06-18 A cost-effective and efficient reprogramming platform for large-scale production of integration-free human induced pluripotent stem cells in chemically defined culture Beers, Jeanette Linask, Kaari L. Chen, Jane A. Siniscalchi, Lauren I. Lin, Yongshun Zheng, Wei Rao, Mahendra Chen, Guokai Sci Rep Article Factors limiting the adoption of iPSC technology include the cost of developing lines and the time period that it takes to characterize and bank them, particularly when integration free, feeder free, and Xeno-free components are used. In this manuscript we describe our optimization procedure that enables a single technician to make 20–40 lines at a time in a 24–96 well format in a reliable and reproducible fashion. Improvements spanned the entire workflow and included using RNA virus, reducing cytotoxicity of reagents, developing improved transfection and freezing efficiencies, modifying the manual colony picking steps, enhancing passaging efficiency and developing early criteria of success. These modifications allowed us to make more than two hundred well-characterized lines per year. Nature Publishing Group 2015-06-11 /pmc/articles/PMC4464084/ /pubmed/26066579 http://dx.doi.org/10.1038/srep11319 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Beers, Jeanette
Linask, Kaari L.
Chen, Jane A.
Siniscalchi, Lauren I.
Lin, Yongshun
Zheng, Wei
Rao, Mahendra
Chen, Guokai
A cost-effective and efficient reprogramming platform for large-scale production of integration-free human induced pluripotent stem cells in chemically defined culture
title A cost-effective and efficient reprogramming platform for large-scale production of integration-free human induced pluripotent stem cells in chemically defined culture
title_full A cost-effective and efficient reprogramming platform for large-scale production of integration-free human induced pluripotent stem cells in chemically defined culture
title_fullStr A cost-effective and efficient reprogramming platform for large-scale production of integration-free human induced pluripotent stem cells in chemically defined culture
title_full_unstemmed A cost-effective and efficient reprogramming platform for large-scale production of integration-free human induced pluripotent stem cells in chemically defined culture
title_short A cost-effective and efficient reprogramming platform for large-scale production of integration-free human induced pluripotent stem cells in chemically defined culture
title_sort cost-effective and efficient reprogramming platform for large-scale production of integration-free human induced pluripotent stem cells in chemically defined culture
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4464084/
https://www.ncbi.nlm.nih.gov/pubmed/26066579
http://dx.doi.org/10.1038/srep11319
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