Production of germline competent chicken cell lines

描述（由申请人提供）： 这项研究的目的是从鸡原始生殖细胞中建立一个细胞系，保留其对生殖系的贡献潜力，目的是开发在鸡蛋中产生药物蛋白质的转基因鸡。 可以培养从血液和性腺获得的原始生殖细胞，并且当注射到14-17期鸡胚胎的血流中时，原始生殖细胞保持其贡献于生殖系的能力。然而，只报道了短时间的混合培养，并且所产生的细胞数量太少，不能用于生产转基因鸡。如果PGCs可以培养和扩增更长的时间以实现转基因的稳定整合，同时保持其在生殖系中定殖的能力，那么它将为生殖系传播提供一种新颖且可靠的途径。 本申请提出从新月体胚芽中获得PGC以获得纯PGC群体，并在各种条件下培养PGC以获得在注射到14-16期（H&H）胚胎的血流中后保留对种系有贡献的潜力的细胞系。

DESCRIPTION (provided by applicant): The aim of this research is to establish a cell line from chicken primordial germ cells that retains it potential to contribute to the germline with the goal of developing genetically modified chickens that produce pharmaceutical proteins in their eggs. Primordial germ cells obtained from the blood and the gonads can be cultured and retain their ability to contribute to the germline when injected into the bloodstream of stage 14-17 chicken embryos. However, only short periods of mixed cultures have been reported and the number of cells that have been generated is too small to be used for the production of transgenic chickens. If PGCs can be cultured and expanded for longer periods of time to achieve stable integration of transgenes while maintaining their competency to colonize the germline, it would provide a novel and reliable route to germline transmission. This application proposes to obtain PGCs from the germinal crescent to obtain a pure population of PGCs and to culture the PGCs under various conditions to obtain a cell line that retains the potential to contribute to the germline after injection into the bloodstream of a stage 14-16 (H&H) embryo.