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Fig. 4 | Stem Cell Research & Therapy

Fig. 4

From: Previously claimed male germline stem cells from porcine testis are actually progenitor Leydig cells

Fig. 4

Induction of sperm differentiation in C1 and C2 clusters. a The C1 were cultured on feeder cells, and they formed Apr spermatogonia (2-cell), Aal-4 spermatogonia (4-cell), and Aal-8 and Aal-16 spermatogonia (8-cell and 16-cell) at days 15, 17, and 20 of culture, respectively. The majority of differentiated spermatogonia were observed at days 25–28 of culture, whereas the sperm cells with long tails were largely observed at day 30 of culture. b Images show the sperm differentiation induction of the C2 at days 1 and 30. c Ploidy analyses were performed by flow cytometry using the C1 and testicular fibroblast cells, induced sperm harvested at day 30 of culture, and live sperm harvested from a healthy boar as positive control. d Ploidy analyses were performed by flow cytometry of the C2 at days 1 and 30, and the testicular fibroblasts. e Expression of the meiosis-associated marker SYCP3 was detected in the induced C1, but not the induced C2. The nuclei were stained with Hoechst 33,342 (blue). f RT-PCR analysis shows expression of sperm-specific markers in the C1 and C2 at the beginning (day 1) and end (day 30) of sperm differentiation induction. g The induced sperm can activate the oocytes to initiate early embryonic development after intracytoplasmic sperm injection (ICSI), and exhibited significant differences compared to the negative group (PBS) and the positive group (mature sperm). The ability of the induced sperm to activate the oocytes was lower than that of mature sperm, with lower efficiency of blastocyst production

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