In vitro culture and characterization of duck primordial germ cells

Abstract

This study aimed to isolate, culture, and characterize duck primordial germ cells (PGCs) and to compare these cells with chicken PGCs. We first cultured Muscovy duck (Cairina moschata) circulating PGCs and gonadal PGCs (gPGCs) in the modified serum-containing medium used to amplify chicken PGCs. gPGCs were found to proliferate better in serum-free chemically defined medium than in serum-containing medium. Thereafter, gPGCs were similarly isolated from 2 other duck breeds, the Pekin duck (Anas platyrhynchos) and the hybrid mule duck (C. moschata × A. platyrhynchos), and amplified for a limited period of time in the chemically defined culture condition, but sufficiently to be characterized and transplanted. Cultured gPGCs of all 3 duck breeds were characterized by Periodic acid-Schiff staining, immunocytochemical staining, and expression analysis of germline-specific and pluripotency genes. Cultured duck gPGCs colonized the gonads after being genetically labeled and injected into recipient embryos. Taken together, these results demonstrate that duck PGCs retain their germline characteristics after being isolated, expanded in vitro, and genetically modified. Further studies are required to establish the optimal conditions for long-term culture of duck PGCs, which may involve supplementing the culture medium with other growth factors or compounds.

Dynamics of the transcriptome during chicken embryo development based on primordial germ cells

“…The motivation for this investigation was to show how quantitative gene expression in PGCs changes during chicken embryonic development. An extensive knowledge on avian PGCs has been generated from in vitro studies [ 17 – 19 ]. In contrast, there is limited information on the regulation of gene expression during the migration of PGCs in vivo.…”

In Vitro Culture of Chicken Circulating and Gonadal Primordial Germ Cells on a Somatic Feeder Layer of Avian Origin

“…They used the same embryonic stem avian complete medium with differing FGF concentrations; moreover, an irradiated STO feeder layer was used only for cPGCs. Their conditions enabled a long-term culture of all stem cell types (over 30–40 days) and did not affect the reactivity to specific markers, but only a higher number of cytoplasmic bridges was observed in the gonadal population [ 29 ].…”

Primordial Germ Cell Specification in Vertebrate Embryos: Phylogenetic Distribution and Conserved Molecular Features of Preformation and Induction

“…Another PGC marker, DAZL RNA, was found to be specifically localized in chicken oocytes and embryonic cleavage furrows, further supporting the conclusion that chicken specify PGCs via maternal inheritance of germline determinants, rather than induction ( Lee et al, 2016 ). Research involving germ cell development in other avian species, such as quail, duck, pheasant, and guinea fowl have primarily been in the context of isolation and in vitro production of PGCs in hopes of generating long-term PGC culture systems and transgenic progeny, rather than elucidating the early mechanistic details of germ plasm aggregation and maintenance ( Reynaud, 1981 ; Ono et al, 1996 ; Kang et al, 2008 ; Wernery et al, 2010 ; van de Lavoir et al, 2012 ; Chen et al, 2019 ). Analysis of PGC specification in vitro , integrated with emerging studies in developing embryos, should provide valuable insight into avian germline specification.…”