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Figure S1. Morphology of amniotic fluid-derived progenitor cells. (A) The appearance and growth of MSC-like cell colonies on the fifth day of culturing. (B) The fibroblast-like AFPC colonies grew to confluence in the first passage culture. (C) AFPC at passage 3 appeared homogeneous. (D) AFPC at passage 12 remained fibroblast-like.

Figure S2. AFPCs from all three patients expressed NANOG, Oct4 and SOX2 mRNA. RT-PCR analyses detected NANOG, Oct4, and SOX2 mRNA in cultured AFPCs. hESCs were used as a positive control, and 293 T cells were used for negative control. β-Actin served as the normalization reference.

Figure S3. Immunocytochemical analyses revealed that most AFPCs expressed the intracellular embryonic marker proteins SOX2 and NANOG, but lacked signal for OCT4.

Figure S4. Immunocytochemical analyses revealed that most AFPCs expressed the embryonic surface markersSSEA4 and TRA-1-60, but notTRA-1-81.

Figure S5. Adipogenesis and osteogenesis of amniotic fluid progenitor cells (magnification 200×). (A) hAFPCs were cultured in adipogenic medium for 3 weeks. (B) Adipogenic differentiation detected by Oil Red O staining of lipid-rich cytoplasmic vacuoles (red). (C) hAFPCs were cultured in osteogenic medium for 3 weeks. (D) Osteogenic differentiation detected by staining for alkaline phosphatase enzyme activity (blue).

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