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克隆、干细胞及辅助生殖 PowerPoint PPT Presentation


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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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Sperm motility

Sperm Motility


Sea urchin fertilization scanning electron microscope

Sea Urchin Fertilization: Scanning Electron Microscope


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Clone cloning

clone/cloning

  • clone

  • cloned animal

  • (animal cloning)


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  • embryonic nuclear transfer

  • (somatic nuclear transfer)

  • (intraspecies nuclear transfer)

  • (interspecies nuclear transfer)


What is cloning are there different types of cloning

What is cloning? Are there different types of cloning?

(1) DNADNA

recombinant DNA technology

or DNA cloning

(2) reproductive cloning

(3) therapeutic cloning


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2

3

4

5

6

7


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Piezo


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  • 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.


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  • The Cloning of Dolly


Cumulina

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


Wakayama t 1998

Wakayama T1998


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(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.


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1999

  • April 1999: First (three) cloned goats;

  • Mira, Mira, and Mira...,

  • Louisiana State University and Tufts University School of Veterinary Medicine,

  • Nature Biotechnology.


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  • August 2000: First cloned Piggies:

  • Roslin Institute / Geron

  • National Institute of Animal Industry in Japan (cute picture!)

  • simultaneously publish in Science and Nature

Onishi A

2000


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Shin t 2002

Shin T2002


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  • 20035434649


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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.


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2002

  • Rabbits born from somatic nuclear transfer

  • Chesn, P. et al.

  • Cloned rabbits produced by nuclear transfer from adult somatic cells.

  • Nat Biotechnol 20, 366-369 (April 2002).


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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.


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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.


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(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, Snuppys clone status had to be rechecked after doubts were cast on the work of his creator,

  • Korean scientist Hwang Woo-suk.


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  • 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.


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2006

  • Cloned ferrets produced by somatic cell nuclear transfer

  • Li and Engelhardt,

  • Dev Biol. 2006 May 15; 293: 439448.

  • 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(


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  • 13


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  • 2000

  • 20003

  • 1999


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  • 2001113116

  • 1998


Chen dy 2003 1998

Chen DY2003 1998


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  • 2005

  • 88113085

  • 2000


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  • 199910%

  • Dominko

  • WhiteArgali16~32645159


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  • 2000LanzaBos gaurus1

  • 2001LoiOvis orientalis1


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1

1alpha 1,3

2

2

3therapeutic cloning


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  • August 7th, 2001: Human cloning conference: a joint panel of the National Academy of Science Committees on Science, Engineering Public Policy and the Board of Life Sciences to discuss the ethical and scientific concerns of human cloning

  • "Despite warnings from scientists, 3 privately-funded US / European groups vowed at the meeting (and on televised coverage of the meetings) to continue their programs on Human Cloning.


The three groups currently attempting to clone humans for reproductive purposes

The three groups currently attempting to clone humans for reproductive purposes

1. Dr. Severino Antinori: of Italy, who became infamous in 1994 when he helped Rosanna Della Corte, a 63-year-old woman give birth. Plans to use the man's genetic material and the woman's egg (and egg cytoplasm, which contains the woman's mitochondria).

2. Dr. Panos Zavos: of Lexington KY, director of the Andrology Institute of America, and the Kentucky Center for Reproductive Medicine.

3. Dr. Brigitte Boisselier: scientific director of Clonaid, "The First Human Cloning Company" . Founded several years ago by Rael, a spiritual leader of the Raelian Movement, who believes that human life is the result of extraterrestrial genetic experiments.


Bad animal cloning where is animal cloning going to end

Bad Animal Cloning?Where is Animal Cloning going to end?


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1.

  • It took 277 attempts to get 1 cloned sheep - Dolly.

  • It took 305 attempts to get Idaho Gem and Utah Pioneer

  • It took 460 attempts to get Kaguya(


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2.

  • (<4%)

  • Large Offspring Syndrome


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3. DNA


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  • stem cell

  • (embryonic stem cell, ES cell)

  • (Embryonic germ cells, EG cells)


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2007

  • 2007:

  • ---

  • DNA


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  • Nature. 1981 Jul 9; 292(5819):154-6.

    Establishment in culture of pluripotential cells from mouse embryos.

    Martin J. Evans-, Kaufman MH

    Department of Genetics, University of Cambridge, UK.


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    • ;


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  • Totipotent stem cell: ,

  • pluripotent stem cell:

  • multipotent stem cell:


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ES

  • SSEA-1(-1

  • OCT4


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  • ES cells (embryonic stem cells)

  • 1981ES

  • 1998ES


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ES

  • James A Thomson et al, 1998

    Wisconsin Regional Primate Research Center, University of Wisconsin

    Embryonic stem cell lines derived from human blastocysts

    Science 282:1145

  • Shamblott MJ, John D Gearhart, 1998

    Derivation of pluripotent stem cells from cultured human primordial germ cells

    PNAS 95:13726-13731


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  • ???


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  • 2004

    2005Science11


Nat cell biol 2006 apr 8 4 384 90 dyce pw wen l li j university of guelph guelph ontario canada

Nat Cell Biol. 2006 Apr;8(4):384-90. Dyce PW, Wen L, Li J.University of Guelph, Guelph, Ontario, Canada.

  • ZP


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/


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  • Shinya Yamanaka

  • Shinya YamanakaRudolf JaenischKonrad Hochedlinger


Induced pluripotent stem cells

Induced pluripotent stem cells

  • iPS cellsiPSCs

  • 2006IPSCs2007


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  • Generating induced pluripotent stem cells.


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iPS


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Personalized Stem Cells One Step Closer to RealityResearchers Create Disease-Specific, Individualized Human Stem CellsABC News Medical UnitJuly 31, 2008

  • Researchers from Harvard and Columbia Universities used skin cells from two patients with Amyotrophic Lateral Sclerosis (ALS) to create stem cells and then reprogrammed them to morph into replacement motor neurons.

  • "It opens doors to making patient-specific stem cell lines," said Dr. Kevin Eggan, principle faculty member at the Harvard Stem Cell Institute and lead author of a study that was released today in the journal Science.

  • "You can use these cells to make the actual cell type for that person's disease."


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Researchers Use Embryonic Stem Cells to Treat DiabetesStudy in Mice Shows Promise; Hurdles RemainBy AUDREY GRAYSONABC News Medical UnitFeb. 20, 2008

  • Doctors may be one step closer to using stem cells to cure diabetes, according to a new study by researchers at the stem cell engineering company Novacell, Inc. in San Diego who report that they managed to convert human embryonic stem cells into insulin-producing cells.

  • The researchers, who reported their findings in the journal Nature Biotechnology, found that when they injected these human cells into diabetic mice, the treatment alleviated diabetes in the rodents.

  • The new technique used by his team will provide doctors with a bulk supply of clean, uncontaminated insulin-secreting cells


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iPS


Stem cells a new path to pluripotency nature 451 858 14 february 2008

Stem Cells:A new path to pluripotency Nature451:858 (14 February 2008)

  • Soon after, in December, George Daley, of Harvard Medical School in Boston, Massachusetts, and his colleagues also demonstrated iPS cells could be generated from a wide variety of adult cells.

  • "For any patient we can use the technique and take a skin biopsy to establish a pluripotent cell," says Daley.

  • One of the most valuable aspects of the iPS-cell technology currently, he says, is the ability to perform disease modelling.

  • And to take advantage of this, Daley and his colleagues are working on generating large numbers of disease specific iPS-cell lines.


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  • iPS cells

  • iPS

  • OCT4SOX2


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  • Yamanaka12120%

  • Yamanaka


Hypothetical strategy for using ips cells in cell based therapies

Hypothetical strategy for using iPS cells in cell-based therapies


Scientists cure sickle cell anemia in mouse model

Scientists Cure Sickle Cell Anemia in Mouse Model


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  • assisted reproduction

  • reproductive medicine


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  • 15%20%

  • Louise Brown1978725

  • 100,000

  • 1988

  • 174


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  • Zheng Mengzhu (left), China's first test-tube baby, now 20 , kisses professor Zhang Lizhu, 87 - who helped her mother fall pregnant via in vitro fertilization - at a ceremony last month in Beijing. Yang Shen

  • Test tube baby boom2008-03-12


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  • 19891990


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  • Intra Uterine Insemination (IUI)

  • Therapeutic Insemination with Donor Sperm (TID)

  • In-Vitro Fertilization (IVF)

  • Gamete IntraFallopian Transfer (GIFT)

  • IntraCytoplasmic Sperm Injection (ICSI)


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(IUI)


Tid tid

(TID)TID


In vitro fertilization ivf

In-vitro fertilization (IVF)

  • IVF


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GIFT


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(ICSI)

  • IVF


Preimplantation genetic diagnosis pgd

Preimplantation genetic diagnosisPGD)


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  • Microsurgical Epididymal Sperm Aspiration (MESA)

  • Testicular Sperm Extraction (TESE)

  • Round Spermatid Injection (ROSI)

  • Sperm Cryopreservation and Thawing

  • Embryo Cryopreservation and Thawing


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  • Sperm donation

  • Oocyte donation

  • Embryo donation

  • Assisted hatching

  • Traditional surrogacy

  • Gestational surrogacy

  • Gender selection


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X Y

Y X Y

X Y X Y

XY

XX

X

Y


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  • April 2004: Mouse created without fatherKaguya"(Japanese folk tale) Mouse born by combining 2 egg nuclei. Kaguya was the ONLY pup of 460 attempts (but healthy and fertile).

  • Key to the success= use of an immature, female egg that lacked key imprinting proteins H19 and Igf2, making the egg more sperm-like".

  • We have shown for the first time that it is possible to obtain a viable adult mouse from two maternal genomes.").

  • Twenty-seven "bimaternal" mice survived to adulthood!!!


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    20

  • 260-270


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  • 200332001


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