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Optimizing the method for generation of integration-free induced pluripotent stem cells from human peripheral blood

BACKGROUND: Generation of induced pluripotent stem cells (iPSCs) from human peripheral blood provides a convenient and low-invasive way to obtain patient-specific iPSCs. The episomal vector is one of the best approaches for reprogramming somatic cells to pluripotent status because of its simplicity...

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Autores principales: Gu, Haihui, Huang, Xia, Xu, Jing, Song, Lili, Liu, Shuping, Zhang, Xiao-bing, Yuan, Weiping, Li, Yanxin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6002980/
https://www.ncbi.nlm.nih.gov/pubmed/29907164
http://dx.doi.org/10.1186/s13287-018-0908-z
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author Gu, Haihui
Huang, Xia
Xu, Jing
Song, Lili
Liu, Shuping
Zhang, Xiao-bing
Yuan, Weiping
Li, Yanxin
author_facet Gu, Haihui
Huang, Xia
Xu, Jing
Song, Lili
Liu, Shuping
Zhang, Xiao-bing
Yuan, Weiping
Li, Yanxin
author_sort Gu, Haihui
collection PubMed
description BACKGROUND: Generation of induced pluripotent stem cells (iPSCs) from human peripheral blood provides a convenient and low-invasive way to obtain patient-specific iPSCs. The episomal vector is one of the best approaches for reprogramming somatic cells to pluripotent status because of its simplicity and affordability. However, the efficiency of episomal vector reprogramming of adult peripheral blood cells is relatively low compared with cord blood and bone marrow cells. METHODS: In the present study, integration-free human iPSCs derived from peripheral blood were established via episomal technology. We optimized mononuclear cell isolation and cultivation, episomal vector promoters, and a combination of transcriptional factors to improve reprogramming efficiency. RESULTS: Here, we improved the generation efficiency of integration-free iPSCs from human peripheral blood mononuclear cells by optimizing the method of isolating mononuclear cells from peripheral blood, by modifying the integration of culture medium, and by adjusting the duration of culture time and the combination of different episomal vectors. CONCLUSIONS: With this optimized protocol, a valuable asset for banking patient-specific iPSCs has been established. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13287-018-0908-z) contains supplementary material, which is available to authorized users.
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spelling pubmed-60029802018-07-06 Optimizing the method for generation of integration-free induced pluripotent stem cells from human peripheral blood Gu, Haihui Huang, Xia Xu, Jing Song, Lili Liu, Shuping Zhang, Xiao-bing Yuan, Weiping Li, Yanxin Stem Cell Res Ther Short Report BACKGROUND: Generation of induced pluripotent stem cells (iPSCs) from human peripheral blood provides a convenient and low-invasive way to obtain patient-specific iPSCs. The episomal vector is one of the best approaches for reprogramming somatic cells to pluripotent status because of its simplicity and affordability. However, the efficiency of episomal vector reprogramming of adult peripheral blood cells is relatively low compared with cord blood and bone marrow cells. METHODS: In the present study, integration-free human iPSCs derived from peripheral blood were established via episomal technology. We optimized mononuclear cell isolation and cultivation, episomal vector promoters, and a combination of transcriptional factors to improve reprogramming efficiency. RESULTS: Here, we improved the generation efficiency of integration-free iPSCs from human peripheral blood mononuclear cells by optimizing the method of isolating mononuclear cells from peripheral blood, by modifying the integration of culture medium, and by adjusting the duration of culture time and the combination of different episomal vectors. CONCLUSIONS: With this optimized protocol, a valuable asset for banking patient-specific iPSCs has been established. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13287-018-0908-z) contains supplementary material, which is available to authorized users. BioMed Central 2018-06-15 /pmc/articles/PMC6002980/ /pubmed/29907164 http://dx.doi.org/10.1186/s13287-018-0908-z Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Short Report
Gu, Haihui
Huang, Xia
Xu, Jing
Song, Lili
Liu, Shuping
Zhang, Xiao-bing
Yuan, Weiping
Li, Yanxin
Optimizing the method for generation of integration-free induced pluripotent stem cells from human peripheral blood
title Optimizing the method for generation of integration-free induced pluripotent stem cells from human peripheral blood
title_full Optimizing the method for generation of integration-free induced pluripotent stem cells from human peripheral blood
title_fullStr Optimizing the method for generation of integration-free induced pluripotent stem cells from human peripheral blood
title_full_unstemmed Optimizing the method for generation of integration-free induced pluripotent stem cells from human peripheral blood
title_short Optimizing the method for generation of integration-free induced pluripotent stem cells from human peripheral blood
title_sort optimizing the method for generation of integration-free induced pluripotent stem cells from human peripheral blood
topic Short Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6002980/
https://www.ncbi.nlm.nih.gov/pubmed/29907164
http://dx.doi.org/10.1186/s13287-018-0908-z
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