Fig. 10: Establishment of Japanese golden eagle iPSCs from somatic cells derived from emerging pinfeather.

a Structure of the reprogramming vectors. b Establishment flow of Japanese golden eagle iPSCs with PB-DDR-8F. c Morphological features of Japanese golden eagle-derived primary and established iPSC colonies. Panels show the primary colonies of reprogrammed cells (i–iii), and established iPSC colonies (iv–vi). Panels show merged images (i, iv), bright field images (ii, v), and GFP fluorescence images (iii, vi). The bars indicate 500 μm (primary colony) and 200 μm (Established iPSCs). d Detection of reprogramming cassettes (PB-DDR-8F vector) and Tsc-2 gene (internal control) using genomic PCR. e DNA content profile analysis of Japanese golden eagle-derived somatic cells and iPSCs (clone1–3) using flow cytometry. f Karyotype analysis of Japanese golden eagle-derived iPSCs. The panel shows the representative mitotic phase of the Japanese golden eagle cells (i), and the aligned chromosomes in the Japanese golden eagle cells (ii). The panel shows the chromosome numbers of the Japanese golden eagle fibroblasts and iPSCs (iii). g Alkaline phosphatase (AP) staining. AP staining of Japanese golden eagle-derived primary colonies (i, ii). The bar indicates 500 µm. AP staining of Japanese golden eagle-derived iPSCs (iii, iv). The bar indicates 500 µm. h Expression of pluripotency markers in iPSCs. Panels show SSEA-1 (i, ii), SSEA-3 (iii, iv), and SSEA-4 (v, vi). Merged images (i, iii, v), SSEA-derived fluorescence images (ii, iv, vi). The bar indicates 200 µm. i Pluripotency-related gene expression in Japanese golden eagle-derived somatic cells and iPSCs (passages 4 and 10). Centerlines of box plots indicate medians; box limits indicate the 25th and 75th percentiles. n = 6. *P < 0.05. j Study summary.