Generation of a human embryonic stem cell line, NERCe003-A-1, with lentivirus vector-mediated inducible CTNNB1 overexpression

Yang Wang; Juan Yu; Lvjun Liu; Wen Li; Xingxiang Duan; Yingying Peng; Sicong Zeng; Qi Ouyang; Guangxiu Lu; Ge Lin; Yi Sun

doi:10.1016/j.scr.2018.01.020

Generation of a human embryonic stem cell line, NERCe003-A-1, with lentivirus vector-mediated inducible CTNNB1 overexpression

Stem Cell Res. 2018 Apr:28:61-65. doi: 10.1016/j.scr.2018.01.020. Epub 2018 Feb 3.

Authors

Yang Wang¹, Juan Yu², Lvjun Liu¹, Wen Li³, Xingxiang Duan¹, Yingying Peng¹, Sicong Zeng⁴, Qi Ouyang⁵, Guangxiu Lu⁶, Ge Lin⁷, Yi Sun⁸

Affiliations

¹ Institute of Reproductive and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha, China.
² National Engineering and Research Center of Human Stem Cells, Changsha, China.
³ Institute of Reproductive and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha, China; Key Laboratory of Stem Cells and Reproductive Engineering, Ministry of Health, Changsha, China; Reproductive & Genetic Hospital of CITIC-Xiangya, Changsha, China.
⁴ Institute of Reproductive and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha, China; National Engineering and Research Center of Human Stem Cells, Changsha, China.
⁵ Institute of Reproductive and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha, China; National Engineering and Research Center of Human Stem Cells, Changsha, China; Key Laboratory of Stem Cells and Reproductive Engineering, Ministry of Health, Changsha, China.
⁶ National Engineering and Research Center of Human Stem Cells, Changsha, China; Key Laboratory of Stem Cells and Reproductive Engineering, Ministry of Health, Changsha, China; Reproductive & Genetic Hospital of CITIC-Xiangya, Changsha, China.
⁷ Institute of Reproductive and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha, China; National Engineering and Research Center of Human Stem Cells, Changsha, China; Key Laboratory of Stem Cells and Reproductive Engineering, Ministry of Health, Changsha, China; Reproductive & Genetic Hospital of CITIC-Xiangya, Changsha, China.
⁸ Institute of Reproductive and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha, China; National Engineering and Research Center of Human Stem Cells, Changsha, China; Key Laboratory of Stem Cells and Reproductive Engineering, Ministry of Health, Changsha, China. Electronic address: [email protected].

PMID: 29428849
DOI: 10.1016/j.scr.2018.01.020

Abstract

The human embryonic stem cell (hESC) line NERCe003-A-1 was generated by introducing lentiviral-vector-mediated tetracycline-inducible β-catenin expression into a normal hESC line, NERCe003-A. The resulting cell line can overexpress the β-catenin protein, encoded by the CTNNB1 gene, after exposure to doxycycline (Dox). CTNNB1 gene expression was confirmed by quantitative PCR (qPCR) and immunofluorescence assays. Further characterization confirmed that the NERCe003-A-1 cell line expresses typical pluripotency markers and has the ability to form the three germ layers both in vitro and in vivo.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Culture Techniques / methods*
Cell Differentiation
Cell Line
Genetic Vectors / metabolism*
Human Embryonic Stem Cells / cytology*
Humans
Karyotyping
Lentivirus / genetics*
Male
Mice
Mycoplasma / isolation & purification
beta Catenin / genetics*
beta Catenin / metabolism

Substances

CTNNB1 protein, human
beta Catenin