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克隆、干细胞及辅助生殖

克隆、干细胞及辅助生殖. 杨增明 厦门大学生命学院. Sperm Motility. Sea Urchin Fertilization: Scanning Electron Microscope. 内容. 克隆 干细胞 辅助生殖. 一、克隆（ clone/cloning ）. “ 克隆 ”一词是来源于英文单词 clone 的音译，其原义是通过无性形式由单个细胞产生的，和亲代非常相象的个体。 克隆动物 （ cloned animal ）是指由一个动物经无性繁殖或孤雌生殖而产生的后代，克隆个体和被克隆个体在遗传上是完全相同的。

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克隆、干细胞及辅助生殖

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  1. 克隆、干细胞及辅助生殖 杨增明 厦门大学生命学院

  2. Sperm Motility

  3. Sea Urchin Fertilization: Scanning Electron Microscope

  4. 内容 • 克隆 • 干细胞 • 辅助生殖

  5. 一、克隆（clone/cloning） • “克隆”一词是来源于英文单词clone的音译，其原义是通过无性形式由单个细胞产生的，和亲代非常相象的个体。 • 克隆动物（cloned animal）是指由一个动物经无性繁殖或孤雌生殖而产生的后代，克隆个体和被克隆个体在遗传上是完全相同的。 • 动物克隆(animal cloning)是指制作克隆动物的方法，主要有胚胎分割和核移植两种方法。

  6. 根据核供体细胞的分化程度，可将核移植分为 • 胚胎细胞核移植（embryonic nuclear transfer） • 体细胞核移植(somatic nuclear transfer)。 根据核供体和核受体是否同一物种，又可分为 • 同种核移植(intraspecies nuclear transfer) • 异种核移植(interspecies nuclear transfer)。

  7. What is cloning? Are there different types of cloning? (1) 重组DNA技术或DNA克隆 （recombinant DNA technology or DNA cloning） (2) 生殖性克隆（reproductive cloning） (3) 治疗性克隆（ therapeutic cloning）

  8. 无性生殖 • 在无性生殖中，一个个体可分成两个或两个以上相同或不同的部分，仅有一个亲本的参与，生殖过程中没有配子的形成。 • 在有性生殖过程中，特化的雄性生殖细胞和雌性生殖细胞发生融合，形成的合子同时携带有两个亲本的遗传信息。

  9. 自然条件下的无性生殖的七种方式 （1）出芽生殖 （2）分裂生殖 （3）断裂 （4）质裂生殖 （5）胞囊生殖 （6）孤雌生殖 （7）孤雄生殖

  10. 有性生殖 • 在有性生殖的生物体（高等生物）中含有两大类细胞， 一是构成组织和器官并执行各种功能的体细胞 二是携带有特定的遗传信息并具有受精后形成合子能力的生殖细胞。 生殖细胞包括卵子和精子两类。有性生殖周期是体细胞与生殖细胞相互转变的过程。在高等生物的机体中，只有一小部分细胞为生殖细胞，然而它们却是正常生命周期中的一个关键环节。

  11. Piezo压电式显微操作仪

  12. Somatic cell nuclear transferInterspecies SCNT. (A) Enucleation of bovine oocyte at metaphase II; (B) Selection of adult human donor cell; (C) Injection of adult human donor cell into the perivitelline space. (D) Human donor cell attached to the enucleated bovine oocyte.

  13. 第一个克隆的哺乳动物 • The Cloning of Dolly

  14. 第一个克隆小鼠（Cumulina） • In 1997, scientists cloned the first mouse from adult cells, calling her Cumulina. • Cumulina died at the age of two years and seven months at a relatively healthy age for a mouse. • Mice remain one of the more difficult species to clone. • Nature 394:369-374. • Teruhiko Wakayama

  15. 小鼠体细胞克隆、再克隆照片（Wakayama T等，1998）

  16. 克隆牛(1998) • George and Charlie, born in 1998, were the first cloned transgenic calves, paving the way for "pharming". • Scientists hope to combine cloning and genetic modification to create animals capable of producing medicinal products. • One of the calves' creators is now president of a company aiming to produce human antibodies.

  17. 克隆山羊（1999） • April 1999: First (three) cloned goats; • Mira, Mira, and Mira..., • Louisiana State University and Tufts University School of Veterinary Medicine, • Nature Biotechnology.

  18. 克隆猪 • August 2000: First cloned Piggies: • Roslin Institute / Geron • National Institute of Animal Industry in Japan (cute picture!) • simultaneously publish in Science and Nature 首例体细胞克隆猪Onishi A等， 2000）

  19. 半乳糖基转移酶

  20. 体细胞克隆猫（引自Shin T等，2002）

  21. 克隆骡 • 2003年5月4日，经过正常的346天妊娠后，在美国爱达荷大学出生一头体重49公斤的小骡子，这是世界上第一头被克隆的马属动物，也是第一头无生育能力的动物被克隆。

  22. 克隆濒危野牛（2001） • Endangered species In 2001, scientists announced the birth of a cloned gaur, a rare wild ox. • Noah, was created from the skin cells of a male gaur that had died eight years previously; but Noah died just 48 hours after delivery. • Cloning endangered species, or even species that are extinct, has been suggested as a method for conserving animal populations.

  23. 克隆兔（2002） • Rabbits born from somatic nuclear transfer • Chesné, P. et al. • Cloned rabbits produced by nuclear transfer from adult somatic cells. • Nat Biotechnol 20, 366-369 (April 2002).

  24. 克隆大鼠（2003） • Ralph the rat Rats are particularly difficult to clone: the first, Ralph, was produced in 2003. • rat eggs "activate" as soon as they leave the ovaries, making it difficult to introduce new genetic material. • A chemical is needed to stabilise the embryo before cloning is possible. • The French announce a new twist on cloning - rats (great models for diabetes and hypertension) have been much more difficult to clone because their oocytes (necessary for SCNT/cloning) speed through meiosis. • Zhou et al succeeding in cloning Ralph (the rat) by using a drug to block the first metaphase-anaphase transition, a technique they call delayed oocyte activation.

  25. 克隆马（2003） • August 2003: World's first cloned horse born to its genetic twin: -- "Italian scientists have created the world's first cloned horse, Prometea, from an adult cell taken from the horse who gave birth to her. ". • Nature 2003 Aug 7;424:635. Pregnancy: a cloned horse born to its dam twin.

  26. 克隆狗(2003) • a, Snuppy, the first cloned dog, at 67 days after birth (right), with the three-year-old male Afghan hound (left) whose somatic skin cells were used to clone him. Snuppy is genetically identical to the donor Afghan hound. • b, Snuppy (left) was implanted as an early embryo into a surrogate mother, the yellow Labrador retriever on the right, and raised by her. • However, in 2006, Snuppy’s clone status had to be rechecked after doubts were cast on the work of his creator, • Korean scientist Hwang Woo-suk（黄禹锡）.

  27. 克隆狼 • Kim MK, Jang G, Oh HJ, Yuda F, Kim HJ, Hwang WS, Hossein MS, Kim JJ, Shin NS, Kang SK, Lee BC.Endangered wolves cloned from adult somatic cells. • Cloning Stem Cells. 2007 Spring;9(1):130-7. • Erratum in: Cloning Stem Cells. 2007 Autumn;9(3):450.

  28. 克隆雪貂（2006） • Cloned ferrets produced by somatic cell nuclear transfer • Li and Engelhardt, • Dev Biol. 2006 May 15; 293: 439–448. • Why? • The domestic ferret (Mustela putorius furo) is the ideal animal model for influenza infections and potentially other human respiratory diseases such as cystic fibrosis(囊性纤维化）

  29. 多少种哺乳动物已被克隆？ • 共有13种 • 绵羊、山羊、牛、小鼠、大鼠、猪、马 骡子、猫、狗、兔、猴、貂、狼、

  30. 中国的克隆动物 • 山羊：西北农林科技大学 • 绵羊：中科院发育所 • 牛：莱阳农学院？中科院动物所？ • 猪：中国农业大学

  31. 左图为2000年在西北农林科技大学出生的第一头山羊，左图为2000年在西北农林科技大学出生的第一头山羊， • 中间为20003年出生的第三代克隆羊 • 国际上1999年首次得到

  32. 于2001年11月3日和11月6日分别做出我国首例和第二例健康成活的体细胞克隆牛“康康”、“双双”于2001年11月3日和11月6日分别做出我国首例和第二例健康成活的体细胞克隆牛“康康”、“双双” • 国际上1998年

  33. 我国首次完成的成年体细胞克隆牛群体（Chen DY等，2003） 国际上1998年

  34. 2005年问世的我国第一头体细胞克隆猪。 • 8月8日，中国农业大学对外宣布，李宁教授领导的课题组经过一年多的科技攻关，一头体重1130克的克隆小香猪于8月5日在河北省三河市明慧养猪公司顺利诞生。这是我国独立自主完成的首例体细胞克隆猪，表明了我国在此项研究上已经达到了国际先进水平。 • 国际上2000年首次得到

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