Pluripotent State Induction in Mouse Embryonic Fibroblast Using mRNAs of Reprogramming Factors
Reprogramming of somatic cells has great potential to provide therapeutic treatments for a number of diseases as well as provide insight into mechanisms underlying early embryonic development. Improvement of induced Pluripotent Stem Cells (iPSCs) generation through mRNA-based methods is currently an...
Main Authors: | , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2014-11-01
|
Series: | International Journal of Molecular Sciences |
Subjects: | |
Online Access: | http://www.mdpi.com/1422-0067/15/12/21840 |
id |
doaj-0cfe7f14039443ae8999fd462d6d07e9 |
---|---|
record_format |
Article |
spelling |
doaj-0cfe7f14039443ae8999fd462d6d07e92020-11-24T21:53:28ZengMDPI AGInternational Journal of Molecular Sciences1422-00672014-11-011512218402186410.3390/ijms151221840ijms151221840Pluripotent State Induction in Mouse Embryonic Fibroblast Using mRNAs of Reprogramming FactorsAhmed Kamel El-Sayed0Zhentao Zhang1Lei Zhang2Zhiyong Liu3Louise C. Abbott4Yani Zhang5Bichun Li6Provincial Key Laboratory of Molecular Design, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, ChinaProvincial Key Laboratory of Molecular Design, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, ChinaProvincial Key Laboratory of Molecular Design, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, ChinaProvincial Key Laboratory of Molecular Design, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, ChinaDepartment of Veterinary Integrative Biosciences (VIBS), College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX 77843-4458, USAProvincial Key Laboratory of Molecular Design, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, ChinaProvincial Key Laboratory of Molecular Design, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, ChinaReprogramming of somatic cells has great potential to provide therapeutic treatments for a number of diseases as well as provide insight into mechanisms underlying early embryonic development. Improvement of induced Pluripotent Stem Cells (iPSCs) generation through mRNA-based methods is currently an area of intense research. This approach provides a number of advantages over previously used methods such as DNA integration and insertional mutagenesis. Using transfection of specifically synthesized mRNAs of various pluripotency factors, we generated iPSCs from mouse embryonic fibroblast (MEF) cells. The genetic, epigenetic and functional properties of the iPSCs were evaluated at different times during the reprogramming process. We successfully introduced synthesized mRNAs, which localized correctly inside the cells and exhibited efficient and stable translation into proteins. Our work demonstrated a robust up-regulation and a gradual promoter de-methylation of the pluripotency markers, including non-transfected factors such as Nanog, SSEA-1 (stage-specific embryonic antigen 1) and Rex-1 (ZFP-42, zinc finger protein 42). Using embryonic stem cells (ESCs) conditions to culture the iPS cells resulted in formation of ES-like colonies after approximately 12 days with only five daily repeated transfections. The colonies were positive for alkaline phosphatase and pluripotency-specific markers associated with ESCs. This study revealed the ability of pluripotency induction and generation of mouse mRNA induced pluripotent stem cells (mRNA iPSCs) using transfection of specifically synthesized mRNAs of various pluripotency factors into mouse embryonic fibroblast (MEF) cells. These generated iPSCs exhibited molecular and functional properties similar to ESCs, which indicate that this method is an efficient and viable alternative to ESCs and can be used for further biological, developmental and therapeutic investigations.http://www.mdpi.com/1422-0067/15/12/21840pluripotencyreprogrammingepigeneticsinduced pluripotent stem cells in vitro transcription |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Ahmed Kamel El-Sayed Zhentao Zhang Lei Zhang Zhiyong Liu Louise C. Abbott Yani Zhang Bichun Li |
spellingShingle |
Ahmed Kamel El-Sayed Zhentao Zhang Lei Zhang Zhiyong Liu Louise C. Abbott Yani Zhang Bichun Li Pluripotent State Induction in Mouse Embryonic Fibroblast Using mRNAs of Reprogramming Factors International Journal of Molecular Sciences pluripotency reprogramming epigenetics induced pluripotent stem cells in vitro transcription |
author_facet |
Ahmed Kamel El-Sayed Zhentao Zhang Lei Zhang Zhiyong Liu Louise C. Abbott Yani Zhang Bichun Li |
author_sort |
Ahmed Kamel El-Sayed |
title |
Pluripotent State Induction in Mouse Embryonic Fibroblast Using mRNAs of Reprogramming Factors |
title_short |
Pluripotent State Induction in Mouse Embryonic Fibroblast Using mRNAs of Reprogramming Factors |
title_full |
Pluripotent State Induction in Mouse Embryonic Fibroblast Using mRNAs of Reprogramming Factors |
title_fullStr |
Pluripotent State Induction in Mouse Embryonic Fibroblast Using mRNAs of Reprogramming Factors |
title_full_unstemmed |
Pluripotent State Induction in Mouse Embryonic Fibroblast Using mRNAs of Reprogramming Factors |
title_sort |
pluripotent state induction in mouse embryonic fibroblast using mrnas of reprogramming factors |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1422-0067 |
publishDate |
2014-11-01 |
description |
Reprogramming of somatic cells has great potential to provide therapeutic treatments for a number of diseases as well as provide insight into mechanisms underlying early embryonic development. Improvement of induced Pluripotent Stem Cells (iPSCs) generation through mRNA-based methods is currently an area of intense research. This approach provides a number of advantages over previously used methods such as DNA integration and insertional mutagenesis. Using transfection of specifically synthesized mRNAs of various pluripotency factors, we generated iPSCs from mouse embryonic fibroblast (MEF) cells. The genetic, epigenetic and functional properties of the iPSCs were evaluated at different times during the reprogramming process. We successfully introduced synthesized mRNAs, which localized correctly inside the cells and exhibited efficient and stable translation into proteins. Our work demonstrated a robust up-regulation and a gradual promoter de-methylation of the pluripotency markers, including non-transfected factors such as Nanog, SSEA-1 (stage-specific embryonic antigen 1) and Rex-1 (ZFP-42, zinc finger protein 42). Using embryonic stem cells (ESCs) conditions to culture the iPS cells resulted in formation of ES-like colonies after approximately 12 days with only five daily repeated transfections. The colonies were positive for alkaline phosphatase and pluripotency-specific markers associated with ESCs. This study revealed the ability of pluripotency induction and generation of mouse mRNA induced pluripotent stem cells (mRNA iPSCs) using transfection of specifically synthesized mRNAs of various pluripotency factors into mouse embryonic fibroblast (MEF) cells. These generated iPSCs exhibited molecular and functional properties similar to ESCs, which indicate that this method is an efficient and viable alternative to ESCs and can be used for further biological, developmental and therapeutic investigations. |
topic |
pluripotency reprogramming epigenetics induced pluripotent stem cells in vitro transcription |
url |
http://www.mdpi.com/1422-0067/15/12/21840 |
work_keys_str_mv |
AT ahmedkamelelsayed pluripotentstateinductioninmouseembryonicfibroblastusingmrnasofreprogrammingfactors AT zhentaozhang pluripotentstateinductioninmouseembryonicfibroblastusingmrnasofreprogrammingfactors AT leizhang pluripotentstateinductioninmouseembryonicfibroblastusingmrnasofreprogrammingfactors AT zhiyongliu pluripotentstateinductioninmouseembryonicfibroblastusingmrnasofreprogrammingfactors AT louisecabbott pluripotentstateinductioninmouseembryonicfibroblastusingmrnasofreprogrammingfactors AT yanizhang pluripotentstateinductioninmouseembryonicfibroblastusingmrnasofreprogrammingfactors AT bichunli pluripotentstateinductioninmouseembryonicfibroblastusingmrnasofreprogrammingfactors |
_version_ |
1725871999701483520 |