Figure 2.

Enhanced Production of Chicken Reprogrammed Stem Cells with OSKMN Combination

(A) CEFs were transfected with four or five transposons expressing the chicken OCT4, SOX2, KLF4, c-MYC genes (OSKM) plus NANOG gene (OSKMN). Clones emerged 10 days after transfection and more alkaline-positive colonies were obtained with the OSKMN combination than with OSKM combination.

(B) Colonies with high, low, and no AP staining display different morphologies. Scale bars, 50 μm.

(C) Transfected CEFs undergo morphological changes 10 days after transfection as revealed by the SSC/FSC profile and start to express SSEA-1 as a typical mESC marker as analyzed by flow cytometry.

(D) The percentages of SSEA-1-positive cells were quantified in two independent transfections of CEF with OSKM and OSKMN combination after 10 and 20 days of transfection revealing the emergence of reprogrammed cells.

(E) Picked colonies from different gene combinations display three different morphotypes: ES-, scale-, and fibroblast-like morphologies. Scale bars, 100 μm.

(F) Among picked colonies, ES-, scale-, and fibroblast-like colony ratio have been determined for each gene combination.