Generation of Birds Maintaining Identical Genome

获得了鸡受精卵。蛋壳在培养皿上裂开了。蛋黄周围的大部分蛋白被去除。一张滤纸被放在一个胚胎周围。滤纸周围的卵膜被外科剪刀剪断。仅获得胚胎层。在显微镜下观察到发育区为透明区或暗区。从鸡胚层中分离出多能细胞。来自胚层(供体)的细胞被注射到受者的男性或女性胚层中，以产生生殖系嵌合体。供体菌株要么是被禁止的普利茅斯岩石(BPR)，要么是丝绸。受体品系为白色来宏犬。供体细胞被注射到男性或女性受体胚胎中。利用体外培养系统对操作后的嵌合胚胎进行培养。根据供体羽毛产生的黑色色素沉着获得体细胞嵌合体。孵化出来的嵌合体一直被饲养到性成熟。根据目前的实验策略，从胚胚层产生了三对雄性和雌性体细胞嵌合体。一对雄性和雌性嵌合体达到了性成熟。当雌雄试配时，产生了一只羽毛中有来自供体品系(BPR)的黑斑的后代。据推测，精子或卵子是从供体细胞产生的。所开发的策略将为未来保持相同基因组的鸟类(与克隆鸟类相似)的产生提供有力的工具。

Fertilized eggs of the chicken were obtained. The egg shells were cracked on Petri dish. Most of the albumen around the egg yolk was removed. A filter paper was put around an embryo. The egg membrane around the filter paper was cut by a surgical scissors. The blastoderm alone was obtained. The developmental region either area pellucida or area opaca was identified under a microscope. Pluripotent cells were isolated from a blastoderm of the chicken. The cells from a blastoderm (donor) were injected into recipients' male or female blasoderms to generate germline chimeras. The donor strains were either Barred Plyomouth Rock (BPR) or Silky. The recipient strain was White Leghorn. The donor cells were injected into either male or female recipient embryos. The manipulated chimeric embryos were cultured by ex vivo culture system. The somatic chimerism were accessed according to the black pigmentation due to the donor feather. The hatched chimeras were raised until sexual maturities. Three pairs of the male and female somatic chimeras derived from a blastoderm were generated according to the present experimental strategies. A pair of male and female chimeras reached sexual maturity. When the male and female were test mated, one offspring which had black spot derived from donor strain (BPR) in the feather was generated. It was suggested that sperm or ova was generated from the donor cells. The developed strategy should be powerful tool for generation of birds maintaining identical genome (similar to clone birds) in the future.