Efficient iPS cell production with the MyoD transactivation domain in serum-free culture.
A major difficulty of producing induced pluripotent stem cells (iPSCs) has been the low efficiency of reprogramming differentiated cells into pluripotent cells. We previously showed that 5% of mouse embryonic fibroblasts (MEFs) were reprogrammed into iPSCs when they were transduced with a fusion gen...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3316619?pdf=render |
| _version_ | 1818764778579951616 |
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| author | Hiroyuki Hirai Nobuko Katoku-Kikyo Peter Karian Meri Firpo Nobuaki Kikyo |
| author_facet | Hiroyuki Hirai Nobuko Katoku-Kikyo Peter Karian Meri Firpo Nobuaki Kikyo |
| author_sort | Hiroyuki Hirai |
| collection | DOAJ |
| description | A major difficulty of producing induced pluripotent stem cells (iPSCs) has been the low efficiency of reprogramming differentiated cells into pluripotent cells. We previously showed that 5% of mouse embryonic fibroblasts (MEFs) were reprogrammed into iPSCs when they were transduced with a fusion gene composed of Oct4 and the transactivation domain of MyoD (called M(3)O), along with Sox2, Klf4 and c-Myc (SKM). In addition, M(3)O facilitated chromatin remodeling of pluripotency genes in the majority of transduced MEFs, including cells that did not become iPSCs. These observations suggested the possibility that more than 5% of cells had acquired the ability to become iPSCs given more favorable culture conditions. Here, we raised the efficiency of making mouse iPSCs with M(3)O-SKM to 26% by culturing transduced cells at low density in serum-free culture medium. In contrast, the efficiency increased from 0.1% to only 2% with the combination of wild-type Oct4 and SKM (OSKM) under the same culture condition. For human iPSCs, M(3)O-SKM achieved 7% efficiency under a similar serum-free culture condition, in comparison to 1% efficiency with OSKM. This study highlights the power of combining the transactivation domain of MyoD with a favorable culture environment. |
| first_indexed | 2024-12-18T08:07:36Z |
| format | Article |
| id | doaj.art-58d0648dfcbe415892e0d31b345fb6ae |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-18T08:07:36Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-58d0648dfcbe415892e0d31b345fb6ae2022-12-21T21:14:58ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0173e3414910.1371/journal.pone.0034149Efficient iPS cell production with the MyoD transactivation domain in serum-free culture.Hiroyuki HiraiNobuko Katoku-KikyoPeter KarianMeri FirpoNobuaki KikyoA major difficulty of producing induced pluripotent stem cells (iPSCs) has been the low efficiency of reprogramming differentiated cells into pluripotent cells. We previously showed that 5% of mouse embryonic fibroblasts (MEFs) were reprogrammed into iPSCs when they were transduced with a fusion gene composed of Oct4 and the transactivation domain of MyoD (called M(3)O), along with Sox2, Klf4 and c-Myc (SKM). In addition, M(3)O facilitated chromatin remodeling of pluripotency genes in the majority of transduced MEFs, including cells that did not become iPSCs. These observations suggested the possibility that more than 5% of cells had acquired the ability to become iPSCs given more favorable culture conditions. Here, we raised the efficiency of making mouse iPSCs with M(3)O-SKM to 26% by culturing transduced cells at low density in serum-free culture medium. In contrast, the efficiency increased from 0.1% to only 2% with the combination of wild-type Oct4 and SKM (OSKM) under the same culture condition. For human iPSCs, M(3)O-SKM achieved 7% efficiency under a similar serum-free culture condition, in comparison to 1% efficiency with OSKM. This study highlights the power of combining the transactivation domain of MyoD with a favorable culture environment.http://europepmc.org/articles/PMC3316619?pdf=render |
| spellingShingle | Hiroyuki Hirai Nobuko Katoku-Kikyo Peter Karian Meri Firpo Nobuaki Kikyo Efficient iPS cell production with the MyoD transactivation domain in serum-free culture. PLoS ONE |
| title | Efficient iPS cell production with the MyoD transactivation domain in serum-free culture. |
| title_full | Efficient iPS cell production with the MyoD transactivation domain in serum-free culture. |
| title_fullStr | Efficient iPS cell production with the MyoD transactivation domain in serum-free culture. |
| title_full_unstemmed | Efficient iPS cell production with the MyoD transactivation domain in serum-free culture. |
| title_short | Efficient iPS cell production with the MyoD transactivation domain in serum-free culture. |
| title_sort | efficient ips cell production with the myod transactivation domain in serum free culture |
| url | http://europepmc.org/articles/PMC3316619?pdf=render |
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