Many studies demonstrated that chicken primordial germ cells (PGCs) could maintain undifferentiated state on mouse embryonic fibroblast feeders supplemented with growth factors and cytokines. However, the xenosupport systems may run risk of cross-transfer of animal pathogens from the other animal feeder, matrix to the PGCs, then influencing later transgenic technology. In this study, chicken PGCs were identified by alkaline phosphatase, stage-specific embryonic antigen-1 and Oct-4 immunocytochemical stainings. Three different homologous somatic cell feeder layers (chicken embryonic fibroblast feeder layer, CEF; embryonic skeletal myoblast feeder layer; follicular granulosa cell feeder layer) were used to support growth and proliferation of PGCs to find a better supporting culture system. In addition, the effects of fetal calf serum (FCS), leukemia inhibitory factor (LIF) and the combination of insulin, transferring and selenite (ITS) on PGC proliferation were compared. Results showed that CEF was the best supporter for PGC growth and proliferation, which was verified by 5-bromo-2'-deoxyuridine incorporation stain. FCS alone or in combination with LIF could significantly promote PGC proliferation in the presence of CEF in ITS medium. This study will contribute to providing a safer supporting system for chicken PGC amplification in vitro, and may be applied in transgenic chicken production and transplantation therapy.
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