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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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
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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. |
format | Online Article Text |
id | pubmed-6002980 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
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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