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Successful Generation of Human Induced Pluripotent Stem Cell Lines from Blood Samples Held at Room Temperature for up to 48 hr

The collection sites of human primary tissue samples and the receiving laboratories, where the human induced pluripotent stem cells (hIPSCs) are derived, are often not on the same site. Thus, the stability of samples prior to derivation constrains the distance between the collection site and the rec...

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Autores principales: Agu, Chukwuma A., Soares, Filipa A.C., Alderton, Alex, Patel, Minal, Ansari, Rizwan, Patel, Sharad, Forrest, Sally, Yang, Fengtang, Lineham, Jonathan, Vallier, Ludovic, Kirton, Christopher M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4624992/
https://www.ncbi.nlm.nih.gov/pubmed/26388286
http://dx.doi.org/10.1016/j.stemcr.2015.08.012
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author Agu, Chukwuma A.
Soares, Filipa A.C.
Alderton, Alex
Patel, Minal
Ansari, Rizwan
Patel, Sharad
Forrest, Sally
Yang, Fengtang
Lineham, Jonathan
Vallier, Ludovic
Kirton, Christopher M.
author_facet Agu, Chukwuma A.
Soares, Filipa A.C.
Alderton, Alex
Patel, Minal
Ansari, Rizwan
Patel, Sharad
Forrest, Sally
Yang, Fengtang
Lineham, Jonathan
Vallier, Ludovic
Kirton, Christopher M.
author_sort Agu, Chukwuma A.
collection PubMed
description The collection sites of human primary tissue samples and the receiving laboratories, where the human induced pluripotent stem cells (hIPSCs) are derived, are often not on the same site. Thus, the stability of samples prior to derivation constrains the distance between the collection site and the receiving laboratory. To investigate sample stability, we collected blood and held it at room temperature for 5, 24, or 48 hr before isolating peripheral blood mononuclear cells (PBMCs) and reprogramming into IPSCs. Additionally, PBMC samples at 5- and 48-hr time points were frozen in liquid nitrogen for 4 months and reprogrammed into IPSCs. hIPSC lines derived from all time points were pluripotent, displayed no marked difference in chromosomal aberration rates, and differentiated into three germ layers. Reprogramming efficiency at 24- and 48-hr time points was 3- and 10-fold lower, respectively, than at 5 hr; the freeze-thaw process of PBMCs resulted in no obvious change in reprogramming efficiency.
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spelling pubmed-46249922015-11-19 Successful Generation of Human Induced Pluripotent Stem Cell Lines from Blood Samples Held at Room Temperature for up to 48 hr Agu, Chukwuma A. Soares, Filipa A.C. Alderton, Alex Patel, Minal Ansari, Rizwan Patel, Sharad Forrest, Sally Yang, Fengtang Lineham, Jonathan Vallier, Ludovic Kirton, Christopher M. Stem Cell Reports Resource The collection sites of human primary tissue samples and the receiving laboratories, where the human induced pluripotent stem cells (hIPSCs) are derived, are often not on the same site. Thus, the stability of samples prior to derivation constrains the distance between the collection site and the receiving laboratory. To investigate sample stability, we collected blood and held it at room temperature for 5, 24, or 48 hr before isolating peripheral blood mononuclear cells (PBMCs) and reprogramming into IPSCs. Additionally, PBMC samples at 5- and 48-hr time points were frozen in liquid nitrogen for 4 months and reprogrammed into IPSCs. hIPSC lines derived from all time points were pluripotent, displayed no marked difference in chromosomal aberration rates, and differentiated into three germ layers. Reprogramming efficiency at 24- and 48-hr time points was 3- and 10-fold lower, respectively, than at 5 hr; the freeze-thaw process of PBMCs resulted in no obvious change in reprogramming efficiency. Elsevier 2015-09-17 /pmc/articles/PMC4624992/ /pubmed/26388286 http://dx.doi.org/10.1016/j.stemcr.2015.08.012 Text en © 2015 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Resource
Agu, Chukwuma A.
Soares, Filipa A.C.
Alderton, Alex
Patel, Minal
Ansari, Rizwan
Patel, Sharad
Forrest, Sally
Yang, Fengtang
Lineham, Jonathan
Vallier, Ludovic
Kirton, Christopher M.
Successful Generation of Human Induced Pluripotent Stem Cell Lines from Blood Samples Held at Room Temperature for up to 48 hr
title Successful Generation of Human Induced Pluripotent Stem Cell Lines from Blood Samples Held at Room Temperature for up to 48 hr
title_full Successful Generation of Human Induced Pluripotent Stem Cell Lines from Blood Samples Held at Room Temperature for up to 48 hr
title_fullStr Successful Generation of Human Induced Pluripotent Stem Cell Lines from Blood Samples Held at Room Temperature for up to 48 hr
title_full_unstemmed Successful Generation of Human Induced Pluripotent Stem Cell Lines from Blood Samples Held at Room Temperature for up to 48 hr
title_short Successful Generation of Human Induced Pluripotent Stem Cell Lines from Blood Samples Held at Room Temperature for up to 48 hr
title_sort successful generation of human induced pluripotent stem cell lines from blood samples held at room temperature for up to 48 hr
topic Resource
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4624992/
https://www.ncbi.nlm.nih.gov/pubmed/26388286
http://dx.doi.org/10.1016/j.stemcr.2015.08.012
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