Studies On The Production Of Transgenic Poultry Via PGCs And Blastodermal Cells

Primordial germ cells (PGCs) and blastodermal cells were employed as intermediates to produce transgenic poultry in this report, ﹐bservations of the primordial germ cells in blood samples from duck and transfection in vivo: Primordial germ cells from the circulating blood of duck embryo at stage 14-22 were observed by histochemical techniques in smear preparation. In the blood smear, the concentration of circulating PGCs was observed to be highest at stage 16 both in sheldrake and mule-duck, and the concentrations are 8.975% and 2.187% respectively; Furthermore liposome solution, consisting of reporter plasmid GFP and In vivo Gene Delivery System, was injected into the marginal veins of sheldrake embryos at stage 15-16. In all embryos reporter gene was observed expressing Gfp among the wing bud, myelencephalon, somite, spinal cord, notochord, splanchnic mesoderm, endoderm, mesonephros, stomach, extraembryonic vessels and ventricular wall. Moreover the GFP signals were detected in the gonad of 3 embryos out of 400. (2) Blastodermal cell as donor in production of transgenic chimeras: Blastodermal cells were disassociated from the sheldrake embryos at stage X and cultured in vitro in DMEM, which was mixed with different concentration of embryonic extracts. We found that the 2-3% of embryonic extracts in DMEM is more suitable for the growth and proliferation of blastodermal cells in vitro; The results of transfecting experiment indicates that the existence of serum does not affect the efficiency of transfection remarkably, but promote the growth of cells; After being cultured and transfected in vitro the donor cells still have the ability tointegrate into the tissues of recipient embryos, including mandibular process,encephalocoele wall, amnion, vitelline vein, ventricle, vitelline plexus, blood cells, wing bud and gonad. But only two out of twenty-four embryos expressed reporter gene in the gonad.