Morphology and characteristics of the established OSCs. (a) Overview of the OSCs immediately after MACS (left) and the established OSCs (right) which formed the typical structure of cell clusters. (b) Reverse transcriptional PCR analysis for expression profile of OSCs and ovarian tissues as the positive control. There were two set of genes: one for oocytes including Gdf9, Nobox, and Zp3 and another one for germ cells including Fragilis, MVH, Prdm1, and Tert, displaying that OSCs are characteristic of germ cells. No RT, PCR of RNA sample without reverse transcription. (c) Immunofluorescence for MVH and Fragilis in established OSCs and STO as negative control. Green, MVH, and Fragilis immunofluorescence; Blue, DAPI. At the bottom are the images (bright field, Green, DAPI, and merge) of STO from left to right. (d) Alkaline phosphatase staining for established OSCs, mESCs, and STO showed that OSCs (arrows) were positive, whereas mESCs were strongly positive. mESCs, mouse embryonic stem cells. (e-f) Cytogenetic analysis for established OSCs showed normal karyotype.