Long-term culture and characterization of goat primordial germ cells.

摘要

This study was performed to characterize goat PGC after culture and cryopreservation. Goat PGC were isolated from day 32 fetuses and cultured on a continuous cell line of murine embryonal fibroblasts (STO) as feeder-cells in the presence of leukaemia inhibitory factor (LIF). The PGC proliferated slowly and showed colony formation in early passages. Frozen-thawed PGC continued to proliferate when stem cell factor (SCF) was added to the culture medium. However, differentiation into epithelial-like polygonal cells or neuronal cells was observed after 1 or 2 passages. The PGC of 1 female and 1 male cell line were characterized by immunocytochemistry. The PGC showed positive staining for anti-stage-specific embryonic antigen-1 (SSEA-1) and FMA-1 (monoclonal antibody produced against a glycoprotein cell surface antigen of the embryonal carcinoma Nulli SCC1), whereas the reactivity to alkaline phosphatase (AP), an established marker for PGC in mice, was inconsistent. After differentiation, PGC lost their positive reaction to SSEA-1, EMA-1 and AP. It is concluded that SSEA-1 and EMA-1 can be used as reliable markers for identifying goat PGC in addition to morphological criteria. Goat PGC can be kept in long-term culture without losing their morphological characteristics and their positive reaction to SSEA-1 and EMA-1, thus providing a promising source of donor-karyoplasts for nuclear transfer procedures.