Fig 1: PCR from amplified bFGF and SDS-PAGE of produced bFGF. A. The expected 459 bp product of bFGF amplified by PCR with primers that added restriction sites at both ends. M; Size marker, P1 and P2 bFGF PCR product bands, PD; bFGF PCR product digested with SacI/EcoRI, VD; pET28 vector digested with SacI/EcoRI, CP-; Colony PCR negative control, CP1-5; Colony PCR results for clones 1 to 5, VUd; Undigested pET28/bFGF vector, VDS; pET28/ bFGF vector digested with SacI, VDS/E and pET28/bFGF vector digested with SacI /EcoRI. B. SDS-PAGE results for different steps of bFGF production. Recombinant his-tagbFGF expressed and purified at a good concentration and purity. M; Protein size marker, L: Lysate, CO; Cut off, W1; Wash 1, W2; Wash 2, W3; Wash 3, E1; Elution 1 and E2; Elution 2.
Fig 2: Teratoma formation by hESC line in the presence of Royan-bFGF for more than six month continues continuous culture. Hematoxylin and eosin staining of paraffin sections of teratomas represent the differentiation of Royan H6 cells into various tissues, including retinal pigment epithelium as a remarkable tissue of ectoderm, cartilage (mesoderm), and intestinal epithelium (endoderm).
Fig 3: Characterization of human embryonic (hESCs) and induced pluripotent stem cells (hiPSCs) expanded in the presence of bFGF. Lines are characterized after 15 to 18 passages. The cell lines retained key properties of pluripotent markers. Morphology of human embryonic (hESCs) and induced pluripotent stem cells (hiPSCs) and expressions of ALP, Oct4, Nanog, SSEA3, TRA-1-60, and TRA-1-81. Nuclei were stained with DAPI (blue). Dot plot diagram of flow cytometry shows co-expression of two markers. The karyotype of hESCs and hiPSCs after several passages with Royan-bFGF was normal.
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