Abstract
BackgroundLong-term maintenance of avian primordial germ cells (PGCs) in vitro has tremendous potential because it can be used to deepen our understanding of the biology of PGCs. A transgenic bioreactor based on the unique migration of PGCs toward the recipients' sex cord via the bloodstream and thereby creating a germline chimeric bird has many potential applications. However, the growth factors and the signaling pathway essential for inducing proliferation of chicken PGCs are unknown.Methodology/Principal FindingsTherefore, we conducted this study to investigate the effects of various combinations of growth factors on the survival and proliferation of PGCs under feeder-free conditions. We observed proliferation of PGCs in media containing bFGF. Subsequent characterization confirmed that the cultured PGCs maintained expression of PGC-specific markers, telomerase activity, normal migrational activity, and germline transmission. We also found that bFGF activates the mitogen-activated protein kinase kinase/extracellular-signal regulated kinase (MEK/ERK) signaling. Also, the expression of 133 transcripts was reversibly altered by bFGF withdrawal.Conclusions/SignificanceOur results demonstrate that chicken PGCs can be maintained in vitro without any differentiation or dedifferentiation in feeder free culture conditions, and subsequent analysis revealed that bFGF is one of the key factors that enable proliferation of chicken PGCs via MEK/ERK signaling regulating downstream genes that may be important for PGC proliferation and survival.
Highlights
Germ cells play important roles in species continuation by delivering genetic information to the generation
Three growth factors, namely leukemia inhibitory factor (LIF), stem cell factor (SCF), and basic fibroblast growth factor (bFGF), and Sandoz inbred mouse-derived thioguanine-resistant and ouabain-resistant (STO) feeder cells are required for the culture of embryonic germ cells (EGCs) [32,33]
Chicken primordial germ cells (PGCs) have been cultured in media supplemented with the same growth factors and feeder cells as mammalian EGCs [19]
Summary
Germ cells play important roles in species continuation by delivering genetic information to the generation. They arise from a small population of cells known as primordial germ cells (PGCs) [1,2,3]. PGCs are initially localized to the central zone of the area pellucida in stage X embryos [4]. They migrate to the germinal crescent at stage 4 (18– 19 h after incubation) [5] and, between stages 10 and 12, move into blood vessels and begin circulating in the bloodstream [6,7]. The growth factors and the signaling pathway essential for inducing proliferation of chicken PGCs are unknown
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