Efficient preparation and labeling of human induced pluripotent stem cells by nanotechnology

Jing Ruan1,*, Jie Shen2,*, Zheng Wang2, Jiajia Ji1, Hua Song1, Kan Wang1, Bin Liu1, Jinhui Li2, Daxiang Cui11Department of Bio-Nano Science and Engineering, Key Laboratory for Thin Film and Microfabrication Technology of the Ministry of Education, National Key Laboratory of Micro/Nano Fabrication Te...

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Autores principales:	Jing Ruan, Jie Shen, Zheng Wang, et al
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2011
Materias:	Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/fd8c5df4ef35466b80d1bbb4b054de8f
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spelling	oai:doaj.org-article:fd8c5df4ef35466b80d1bbb4b054de8f2021-12-02T07:47:00ZEfficient preparation and labeling of human induced pluripotent stem cells by nanotechnology1176-91141178-2013https://doaj.org/article/fd8c5df4ef35466b80d1bbb4b054de8f2011-02-01T00:00:00Zhttp://www.dovepress.com/efficient-preparation-and-labeling-of-human-induced-pluripotent-stem-c-a6413https://doaj.org/toc/1176-9114https://doaj.org/toc/1178-2013Jing Ruan1,, Jie Shen2,, Zheng Wang2, Jiajia Ji1, Hua Song1, Kan Wang1, Bin Liu1, Jinhui Li2, Daxiang Cui11Department of Bio-Nano Science and Engineering, Key Laboratory for Thin Film and Microfabrication Technology of the Ministry of Education, National Key Laboratory of Micro/Nano Fabrication Technology, Research Institute of Micro/Nano Science and Technology, Shanghai Jiao Tong University, Shanghai, People&rsquo;s Republic of China; 2Shanghai Institute of Digestive Diseases, Shanghai Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People&rsquo;s Republic of China. *These two authors contributed equally to this workAbstract: Efficient preparation and labeling of human induced pluripotent stem (iPS) cells is a great challenge in stem cell research and development. With the aim of investigating the feasibility of using nanotechnology to enhance the preparation efficiency of iPS cells and to label iPS cells for long-term tracing and imaging, in this paper, four transcription factor genes, ie, Oct4, Sox2, LIN28, and Nanog, and packaging plasmids such as PSPAX2 and PMD2.G were cotransfected into 293T cells using Generation 5.0 polyamidoamine dendrimer-modified magnetic nanoparticles (dMNPs) as a delivery system. The resultant supernatant liquids were incubated with human fibroblast cells at 37&deg;C for 21 days, then the embryonic stem (ES) cell-like clones were screened, cultured, and identified. Finally, the prepared iPS cells were labeled with fluorescent magnetic nanoparticles (FMNPs). The results showed that dMNPs can efficiently deliver all vectors into 293T cells. The resultant lentiviruses&rsquo; titers were 10-fold more than those based on Lipofectamine&trade; 2000. Reverse transcription polymerase chain reaction analysis showed that four genes (Oct4, Sox2, LIN28, and Nanog) exhibited different expressions in iPS cells. Immunostaining analysis showed that specific surface markers of ES cells such as SSEA-3, SSEA-4, Tra-1-60, and Tra-1-81 were positive in iPS cells, and the terotomas were formed in NOD-SCID mice that were implanted with iPS cells. Red fluorescent signals could be observed in iPS cells labeled with FMNPs by fluorescent microscopy, and the magnetic signals were detected in labeled iPS cells by magnetic resonance imaging. In conclusion, human iPS cells can be efficiently generated using polyamidoamine dMNPs and lentivirus and labeled with FMNPs for long-term observation and tracking, which has great potential application in the research and development of stem cells in the near future.Keywords: induced pluripotent stem cells, polyamidoamine dendrimer-modified magnetic nanoparticles, fluorescent magnetic nanoparticle, preparation, label Jing RuanJie ShenZheng Wanget alDove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2011, Iss default, Pp 425-435 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine (General) R5-920
spellingShingle	Medicine (General) R5-920 Jing Ruan Jie Shen Zheng Wang et al Efficient preparation and labeling of human induced pluripotent stem cells by nanotechnology
description	Jing Ruan1,, Jie Shen2,, Zheng Wang2, Jiajia Ji1, Hua Song1, Kan Wang1, Bin Liu1, Jinhui Li2, Daxiang Cui11Department of Bio-Nano Science and Engineering, Key Laboratory for Thin Film and Microfabrication Technology of the Ministry of Education, National Key Laboratory of Micro/Nano Fabrication Technology, Research Institute of Micro/Nano Science and Technology, Shanghai Jiao Tong University, Shanghai, People&rsquo;s Republic of China; 2Shanghai Institute of Digestive Diseases, Shanghai Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People&rsquo;s Republic of China. *These two authors contributed equally to this workAbstract: Efficient preparation and labeling of human induced pluripotent stem (iPS) cells is a great challenge in stem cell research and development. With the aim of investigating the feasibility of using nanotechnology to enhance the preparation efficiency of iPS cells and to label iPS cells for long-term tracing and imaging, in this paper, four transcription factor genes, ie, Oct4, Sox2, LIN28, and Nanog, and packaging plasmids such as PSPAX2 and PMD2.G were cotransfected into 293T cells using Generation 5.0 polyamidoamine dendrimer-modified magnetic nanoparticles (dMNPs) as a delivery system. The resultant supernatant liquids were incubated with human fibroblast cells at 37&deg;C for 21 days, then the embryonic stem (ES) cell-like clones were screened, cultured, and identified. Finally, the prepared iPS cells were labeled with fluorescent magnetic nanoparticles (FMNPs). The results showed that dMNPs can efficiently deliver all vectors into 293T cells. The resultant lentiviruses&rsquo; titers were 10-fold more than those based on Lipofectamine&trade; 2000. Reverse transcription polymerase chain reaction analysis showed that four genes (Oct4, Sox2, LIN28, and Nanog) exhibited different expressions in iPS cells. Immunostaining analysis showed that specific surface markers of ES cells such as SSEA-3, SSEA-4, Tra-1-60, and Tra-1-81 were positive in iPS cells, and the terotomas were formed in NOD-SCID mice that were implanted with iPS cells. Red fluorescent signals could be observed in iPS cells labeled with FMNPs by fluorescent microscopy, and the magnetic signals were detected in labeled iPS cells by magnetic resonance imaging. In conclusion, human iPS cells can be efficiently generated using polyamidoamine dMNPs and lentivirus and labeled with FMNPs for long-term observation and tracking, which has great potential application in the research and development of stem cells in the near future.Keywords: induced pluripotent stem cells, polyamidoamine dendrimer-modified magnetic nanoparticles, fluorescent magnetic nanoparticle, preparation, label
format	article
author	Jing Ruan Jie Shen Zheng Wang et al
author_facet	Jing Ruan Jie Shen Zheng Wang et al
author_sort	Jing Ruan
title	Efficient preparation and labeling of human induced pluripotent stem cells by nanotechnology
title_short	Efficient preparation and labeling of human induced pluripotent stem cells by nanotechnology
title_full	Efficient preparation and labeling of human induced pluripotent stem cells by nanotechnology
title_fullStr	Efficient preparation and labeling of human induced pluripotent stem cells by nanotechnology
title_full_unstemmed	Efficient preparation and labeling of human induced pluripotent stem cells by nanotechnology
title_sort	efficient preparation and labeling of human induced pluripotent stem cells by nanotechnology
publisher	Dove Medical Press
publishDate	2011
url	https://doaj.org/article/fd8c5df4ef35466b80d1bbb4b054de8f
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_version_	1718399197407870976

Efficient preparation and labeling of human induced pluripotent stem cells by nanotechnology

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