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METHOD FOR ISOLATING GONADAL PRIMORDIAL GERM CELLS AND METHOD FOR PREPARING GERMLINE CHIMERIC AVES

机译:分离生殖腺原始生殖细胞的方法和生殖胚嵌合Aves的制备方法

摘要

PURPOSE: A method for isolating gonadal primordial germ cells and a method for preparing germline chimeric aves are provided, thereby improving purity and isolation yield of gonadal primordial germ cells, and improving preparation efficiency of germline chimeric aves. CONSTITUTION: The method for isolating gonadal primordial germ cells comprises the steps of: (a) treating gonadal primordial germ tissue cells isolated from embryos of aves with a primordial germ cell-specific first antibody; (b) treating the antibody-bound gonadal primordial germ tissue cells with microbeads bound with a second antibody which has magnetism and binds with the first antibody; and (c) cleansing the resulting product of step (b) and separating the gonadal primordial germ cell-second antibody-microbead complex. The method for preparing germline chimeric aves comprises the steps of: (a) treating gonadal primordial germ tissue cells isolated from embryos of aves with a primordial germ cell-specific first antibody; (b) treating the antibody-bound gonadal primordial germ tissue cells with microbeads bound with a second antibody which has magnetism and binds with the first antibody; (c) cleansing the resulting product of step (b) and separating the gonadal primordial germ cell-second antibody-microbead complex; (d) introducing the isolated gonadal primordial germ cells into the embryo of a recipient egg; and (e) culturing and hatching the egg.
机译:目的:提供一种分离性腺原始生殖细胞的方法和一种制备生殖系嵌合aves的方法,从而提高性腺原始生殖细胞的纯度和分离产率,并提高生殖系嵌合aves的制备效率。组成:分离性腺原始生殖细胞的方法包括以下步骤:(a)用原始生殖细胞特异性第一抗体处理从aves胚胎分离的性腺原始生殖组织细胞; (b)用结合有磁性并结合第一抗体的第二抗体的微珠处理结合抗体的性腺原始生殖细胞。 (c)清洗步骤(b)的所得产物并分离性腺原始生殖细胞-第二抗体-微珠复合物。制备种系嵌合aves的方法包括以下步骤:(a)用原始生殖细胞特异性的第一抗体处理从aves的胚中分离出的性腺原始生殖组织细胞; (b)用结合有磁性并结合第一抗体的第二抗体的微珠处理结合抗体的性腺原始生殖细胞。 (c)清洗步骤(b)的所得产物并分离性腺原始生殖细胞-第二抗体-微珠复合物; (d)将分离的性腺原始生殖细胞引入受体卵的胚胎中; (e)培养和孵化卵。

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