Preparation of induced pluripotent stem cells on dishes grafted on oligopeptide under feeder-free conditions
Human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) have potentially therapeutic applications in the treatment of many diseases, due to their unique ability to differentiate into any type of somatic cell. However, the clinical potential of hESCs and hiPSCs is restric...
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my.upm.eprints.350142015-12-28T08:49:17Z http://psasir.upm.edu.my/id/eprint/35014/ Preparation of induced pluripotent stem cells on dishes grafted on oligopeptide under feeder-free conditions Higuchi, Akon Lin, Feng Ling Cheng, Yu Kai Kao, Ta Chun Kumar, S. Suresh Ling, Qing Dong Hou, Chun Han Chen, Da Chung Hsu, Shih Tien Wu, Gwo Jang Human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) have potentially therapeutic applications in the treatment of many diseases, due to their unique ability to differentiate into any type of somatic cell. However, the clinical potential of hESCs and hiPSCs is restricted by the use of mouse embryonic fibroblasts (MEFs) as a feeder layer for these cells. We report that hiPSCs can be successfully generated without the use of a feeder layer of MEFs. We generated hiPSCs by transducing human adipose-derived stem cells (hADSCs) with a retrovirus containing pluripotency genes, and the hiPSCs were cultured on synthetic dishes grafted with an oligopeptide derived from vitronectin (VN-dish). On the fourth day after transduction, the hADSCs transduced with pluripotency genes were transferred to a MEF layer for culturing as a control condition or to VN-dishes for culture. The hiPSC colonies in the MEF-cultures were clearly observed at day 14 after transduction, whereas hiPSC colonies were detected on the VN-dishes after the cells were passaged. When 105 hADSCs were seeded on the dishes, the number of colonies generated on the MEFs was 120 ± 28, while the number of colonies generated on VN-dishes was 25 ± 8. Thus, the efficiency of hiPSC generation on the VN-dishes under feeder-free conditions was lower than hiPSCs cultured on MEFs. However, the hiPSC colonies from VN-dishes demonstrated alkaline phosphatase activity, and immunohistochemistry suggested that the hiPSCs generated on VN-dishes expressed the pluripotency protein, stage-specific embryonic antigen-4 (SSEA-4), under feeder-free conditions. Elsevier 2014-03 Article PeerReviewed Higuchi, Akon and Lin, Feng Ling and Cheng, Yu Kai and Kao, Ta Chun and Kumar, S. Suresh and Ling, Qing Dong and Hou, Chun Han and Chen, Da Chung and Hsu, Shih Tien and Wu, Gwo Jang (2014) Preparation of induced pluripotent stem cells on dishes grafted on oligopeptide under feeder-free conditions. Journal of the Taiwan Institute of Chemical Engineers, 45 (2). 295-301. ISSN 1876-1070; ESSN: 1876-1089 http://www.sciencedirect.com/science/article/pii/S1876107013001685 10.1016/j.jtice.2013.06.022 |
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Human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) have potentially therapeutic applications in the treatment of many diseases, due to their unique ability to differentiate into any type of somatic cell. However, the clinical potential of hESCs and hiPSCs is restricted by the use of mouse embryonic fibroblasts (MEFs) as a feeder layer for these cells. We report that hiPSCs can be successfully generated without the use of a feeder layer of MEFs. We generated hiPSCs by transducing human adipose-derived stem cells (hADSCs) with a retrovirus containing pluripotency genes, and the hiPSCs were cultured on synthetic dishes grafted with an oligopeptide derived from vitronectin (VN-dish). On the fourth day after transduction, the hADSCs transduced with pluripotency genes were transferred to a MEF layer for culturing as a control condition or to VN-dishes for culture. The hiPSC colonies in the MEF-cultures were clearly observed at day 14 after transduction, whereas hiPSC colonies were detected on the VN-dishes after the cells were passaged. When 105 hADSCs were seeded on the dishes, the number of colonies generated on the MEFs was 120 ± 28, while the number of colonies generated on VN-dishes was 25 ± 8. Thus, the efficiency of hiPSC generation on the VN-dishes under feeder-free conditions was lower than hiPSCs cultured on MEFs. However, the hiPSC colonies from VN-dishes demonstrated alkaline phosphatase activity, and immunohistochemistry suggested that the hiPSCs generated on VN-dishes expressed the pluripotency protein, stage-specific embryonic antigen-4 (SSEA-4), under feeder-free conditions. |
format |
Article |
author |
Higuchi, Akon Lin, Feng Ling Cheng, Yu Kai Kao, Ta Chun Kumar, S. Suresh Ling, Qing Dong Hou, Chun Han Chen, Da Chung Hsu, Shih Tien Wu, Gwo Jang |
spellingShingle |
Higuchi, Akon Lin, Feng Ling Cheng, Yu Kai Kao, Ta Chun Kumar, S. Suresh Ling, Qing Dong Hou, Chun Han Chen, Da Chung Hsu, Shih Tien Wu, Gwo Jang Preparation of induced pluripotent stem cells on dishes grafted on oligopeptide under feeder-free conditions |
author_facet |
Higuchi, Akon Lin, Feng Ling Cheng, Yu Kai Kao, Ta Chun Kumar, S. Suresh Ling, Qing Dong Hou, Chun Han Chen, Da Chung Hsu, Shih Tien Wu, Gwo Jang |
author_sort |
Higuchi, Akon |
title |
Preparation of induced pluripotent stem cells on dishes grafted on oligopeptide under feeder-free conditions |
title_short |
Preparation of induced pluripotent stem cells on dishes grafted on oligopeptide under feeder-free conditions |
title_full |
Preparation of induced pluripotent stem cells on dishes grafted on oligopeptide under feeder-free conditions |
title_fullStr |
Preparation of induced pluripotent stem cells on dishes grafted on oligopeptide under feeder-free conditions |
title_full_unstemmed |
Preparation of induced pluripotent stem cells on dishes grafted on oligopeptide under feeder-free conditions |
title_sort |
preparation of induced pluripotent stem cells on dishes grafted on oligopeptide under feeder-free conditions |
publisher |
Elsevier |
publishDate |
2014 |
url |
http://psasir.upm.edu.my/id/eprint/35014/ http://www.sciencedirect.com/science/article/pii/S1876107013001685 |
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1643831327419531264 |
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13.18916 |