Generation of a human embryonic stem cell line stably expressing high levels of the fluorescent protein mCherry

Figure 3 Characterization of the MEL2-mCherry human embryonic stem cell line. (A) Brightfield and (B) red mCherry fluorescence of a colony of MEL2 human embryonic stem cell (hESC) transfected with Ef1a-mCherry plasmid 12 d after Blasticidin selection (scale bar = 100 μm). (C) Brightfield and (D) red mCherry fluorescence of a colony of single-cell passaging-adapted MEL2-mCherry hESC grown on Matrigel (scale bar = 100 μm). (E) POU5F1/OCT4 expression and (F) corresponding DAPI nuclear staining of MEL2-mCherry hESC. (G) NANOG protein expression and (H) corresponding DAPI nuclear staining in nuclei of MEL2-mCherry cells. (I) Normal human female karyotype of MEL2-mCherry hESC at passage 15 (Giemsa stain of a representative metaphase chromosome spread shown). (J) FACS analysis showing high levels of mCherry fluorescence (95.7%, i.e.,) and the presence of high levels of pluripotency-associated surface antigens Tra1-60, SSEA4 and TG30(CD9). Control-a representative primary isotype control antibody staining.