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.
NERCe002-A-3 cells were generated from the normal human embryonic stem cell line NERCe002-A. NERCe002-A-3 cells overexpressed 14-3-3ζ after exposure to doxycycline. 14-3-3ζ protein have the ability to bind a multitude of functionally diverse signalling proteins. The NERCe002-A-3 cell line is considered a model for functional studies of the 14-3-3ζ protein in hESC self-renewal and cell differentiation. Doxycycline-treated NERCe002-A-3 cells showed a>27-fold increase in relative expression of 14-3-3ζ as compared with un-induced cells. Characterization assays proved that NERCe002-A-3 cells express typical markers of pluripotency and have the ability to form the three germ layers in vivo.
The human embryonic stem cell (hESC) line NERCe002-A-1 was generated through lentiviral transduction of the original NERCe002-A-1 hESC line with Zoanthus sp. green fluorescent protein (ZsGreen). Cells that expressed ZsGreen showed a >8.6-fold increase in fluorescence intensity compared with that of cells that expressed enhanced green fluorescent protein. The fluorescent hESC line can aid in identification of biological characteristics in vitro and in vivo by tracking cell growth, migration, and differentiation. Characteristic tests confirmed that the NERCe002-A-1 cell line expressed typical markers of pluripotency and had the capability to form the three germ layers in vivo.
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