Abstract
Long-term maintenance of chicken primordial germ cells (PGCs) in vitro has tremendous potential for transgenic chicken production. Feeder cells are essential for the establishment and culture of chicken PGCs in vitro. Buffalo rat liver (BRL) cells are the most commonly used feeder cells for PGCs culture; however, this feeder layers from other animal species usually cause immunogenic contaminations, compromising the potential of PGCs in applications. Therefore, we tested chicken source mensenchymal stem cell (MSCs) derived from bone marrow as feeder cells to further improve PGC culture conditions. MSCs derived from chicken bone marrow have a powerful capacity to proliferate and secrete cytokines. We found chicken primordial germ cells derived from circulating blood (cPGCs) and gonads (gPGCs) can be maintained and proliferated with MSCs feeder layer cells. PGCs co-cultured on MSCs feeder retained their pluripotency, expressed PGCs specific genes and stemness markers, and maintained undifferentiated state. Our study indicated that the xeno-free MSCs-feeders culture system is a good candidate for growth and expansion of PGCs as the stepping stone for transgenic chicken research.
Highlights
primordial germ cells (PGCs) are progenitors of germ cells and play an important role in the production of early embryonic germ cells [1] [2]
We found chicken primordial germ cells derived from circulating blood and gonads can be maintained and proliferated with mensenchymal stem cell (MSCs) feeder layer cells
Our study indicated that the xeno-free MSCs-feeders culture system is a good candidate for growth and expansion of PGCs as the stepping stone for transgenic chicken research
Summary
PGCs are progenitors of germ cells and play an important role in the production of early embryonic germ cells [1] [2]. For sustained maintenance in vitro, PGCs isolated from circulating blood at stages 14 - 17 and gonadal ridge at stage 28 could be cultured on a feeder layer of Buffalo rat liver (BRL) cells, Mouse Embryonic Fibroblast (MEF) cells, or Sandoz inbred mouse-derived thioguanine-resistant and ouabain-resistant (STO) cells with basic fibroblast growth factor (bFGF) and stem cell factor (SCF) supplementation [5] [6] [7] [8]. Those culture systems use xeno-animal cells as a feeder layer, which may carry the risk of a cross-transfer of pathogens from other animals. The limited survival time, fewer passage number, and slower proliferation rate of the CEF-feeder increased uncertainty and instability in the process of cultivating the PGCs [10]
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