Fig. 2: Characterization of cultured PGCs. | Communications Biology

Fig. 2: Characterization of cultured PGCs.

From: PGC-based cryobanking, regeneration through germline chimera mating, and CRISPR/Cas9-mediated TYRP1 modification in indigenous Chinese chickens

Fig. 2

A Morphologic characteristics of cultured PGCs. Cultured PGCs showed distinct features, including large nuclei and the presence of abundant lipids in their cytoplasm. Scale bars represent 20 μm. B Immunohistochemistry for pan-germ cell markers in PGC lines. (a) nuclear staining (DAPI), (b) SSEA-1, (c) merged image of SSEA-1 and DAPI. SSEA-1 was predominantly localized to the cell membrane. Scale bars represent 50 µm. C RT-PCR analysis of germ cell-related genes in cultured PGCs. RT-PCR was performed on cDNA samples from cultured PGCs to demonstrate the expression of pan-germ cell-related genes, CVH and DAZL. CEF, chicken embryonic fibroblasts; NC, negative control (H2O). D, G Gonadal migration assay of cultured PGCs into recipient gonads. Cultured PGCs (D) were transiently fluorescent labeled with PKH26 and transplanted into HH14–16 recipient embryos. Scale bars represent 50 μm. Manipulated embryos were cultured in host eggshells until day 7–8 (HH30–32), and recipient gonads, shown as yellow dashed circles, were collected (E). The female recipient embryo is on the left and the male recipient embryo is on the right. The PKH26-labeled PGCs (red cells) showed successful migration into the recipient gonads (F–G). D (left), E, F (left) and G (left) were photographed under bright light, while panels (D) (right), (F) (right) and (G) (right) were photographed under UV-light. The fluorescently labeled PGCs (red cells) were observed in the gonads of both the male and female recipient embryos. Scale bars represent 100 μm.

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