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Establishment of avian biodiversity preservation platform via primordial germ cells (PGCs)
germplasm preservation,chicken,duck,primordial germ cell (PGC),three-dimensional (3D) culture,hybrid sterility,
|Publication Year :||2018|
Primordial germ cells (PGCs) are the progenitors of gametes and could be seen as the key elements for germplasm preservation and the management of avian genetic biodiversity. To obtain the gametes and offspring, a recipient with no endogenous germ cells would be ideal to improve the germline transmission of the donor transplanted PGCs. In duck species, such bird exists as represented by the mule duck, produced by the intergeneric crossing between Muscovy drake (Cairina moschata) and female domestic duck (Anas platyrhynchos). This hybrid has normal somatic development but is impaired in the gamete production leading to hybrid sterility. Therefore, we aim to investigate the cause of this sterility and ultimately to test such breed as the universal recipient for transfer of exogenous duck PGCs to produce gametes of the donor genotype.
The first aim of the present project was to isolate and establish an in vitro culture of duck PGCs derived and adapted from the well-known chicken culture system. First, duck PGCs were successfully isolated from Muscovy, Pekin and hybrid mule ducks. Their germline characteristics and developmental potential were analyzed and were shown to be retained after being expanded in vitro and genetically modified. Second, those PGCs were transplanted to test the gametogenesis in mule duck but no sperm or ovum was produced in transplanted mule ducks. Instead, a large number of cells and debris was found in the semen collected from the mule drakes with PGC transplantation. Among others, mRNA levels of the gonadal somatic enzymes for retinoic acid (RA) synthesis and degradation showed asynchronous dynamic patterns in different developmental stages among parental species and mule ducks. Those observations could lead to the hypothesis of an impairment of the germ cell differentiation in the mule duck gonad that could be due to an altered germ-somatic cell-cell interaction leading to the sterility of the transplanted mule duck.
The second aim of the project was to develop a process of large-scale amplification of avian germ cells. For that, a chemically defined medium was established to grow and amplify chicken PGCs in long-term culture. By adding a functional polymer in the medium, a three-dimensional (3D) suspension culture without mechanical stirring was established leading to a scalable production of PGCs. This system could also contribute to germplasm preservation by amplifying the genetic resource as PGCs of avian species in both genders. Those large-scale produced cells could thus be easily cryopreserved. Additionally, avian specific glycosylation has attracted therapeutic interests and the defined 3D suspension culture system could provide a useful platform for large-scale production of avian PGCs to produce recombinant proteins and vaccines.
|Appears in Collections:||生物科技研究所|
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