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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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Main Authors: 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
Format: Article
Published: Elsevier 2014
Online Access:http://psasir.upm.edu.my/id/eprint/35014/
http://www.sciencedirect.com/science/article/pii/S1876107013001685
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spelling 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
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
description 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
_version_ 1643831327419531264
score 13.211869

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