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The Development of New Technologies in Human Reproduction

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Introduction Human reproductive cloning today continues to preoccupy the general public and its critics in a very controversial manner. – PowerPoint PPT presentation

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Title: The Development of New Technologies in Human Reproduction


1
TO CLONE OR NOT TO CLONE? WHOSE CHOICE IS IT
ANYWAY?
Professor Dr. Panos Zavos, Ed.S., Ph.D.
Professor Emeritus of Reproductive
Physiology/Andrology, University of Kentucky
Director, Andrology Institute of
America Associate Director, Kentucky Center for
Reproductive Medicine IVF, Lexington, KY,
USA Executive Director, The Zavos Organization
2
Introduction
  • Human reproductive cloning today continues to
    preoccupy the general public and its critics in a
    very controversial manner. There is also some
    public hostility directed against it.

3
Public hostility to human reproductive cloning
may be based on an illogical transient fear of a
new technology
The British Medical Association
4
Infertility is a disease
Today, infertility is a disease that reaches
epidemic proportions throughout the developing
World
5
In Vitro Fertilization (IVF)
  • Low sperm count and/or motility
  • Variety of female factors
  • Success rate 33 live birth/transfer

Did not overcome severe asthenospermia
6
Intra Cytoplasmic Sperm Injection (ICSI)
  • If patient has low sperm count or having sperm
    with no motility
  • Success rate 32 live birth/transfer

Required presence of sperm in the ejaculate
7
Sperm and Oocyte Donation
  • No mature sperm present
  • No oocyte (egg) production

Offspring not genetically or biologically related
8
Quotes from childless patients
  • ..we want a child (yesterday, if possible) and a
    healthy child.
  • ..do not want to have another persons spermor
    eggs.
  • ..what other options do we have?
  • we want to have a biological child of our own.

(Received via e-mail from patients)
9
How Therapeutic Cloning Works
  • First, the nucleus of a donor egg is removed.
  • Then a whole somatic cell or the nucleus of a
    whole somatic cell from a patient is inserted.
  • The result is an egg with the patient's genetic
    material.
  • The egg is then induced (jump start) to divide
    and become an embryo which grows into several
    stem cells, all of which are genetically
    identical to the donor cell.

10
Sexual and Asexual Reproduction
No difference in the type of oocyte (egg) used
11
Sexual and Asexual Reproduction
  • Fusion of male and female genetic material
    (pronuclei)

Electrofusion of somatic cell that carries ONLY
the male of female genetic material to the
oocyte
12
Sexual and Asexual Reproduction
No difference in the cell division stages after
fertilization
13
Cloning in Animals
  • Various species have been used as biological
    models for this effort but extensive research on
    somatic cell nuclear transfer (SCNT) has been
    performed using the bovine model.

14
Our Studies
  • In our studies we set out to examine the ability
    of the bovine metaphase oocyte cytoplasm to
    support mitotic cell cycles under the direction
    of differentiated somatic cell nuclei of human
    granulosa cells and fibroblast cells in order to
    test the efficiency of our SCNT techniques.
  • Hybrid embryos

15
Materials and Methods
  • Bovine oocytes were randomly treated either for
    induction of parthenogenesis or for enucleation
    and SCNT using either human granulosa cells or
    human fibroblast cells.

16
Materials and Methods
  • Bovine oocytes were enucleated by aspiration of
    the first polar body and the metaphase plate.

17
Materials and Methods
  • Human granulosa cells and fibroblast cells were
    aspirated into a micropipette.

18
Materials and Methods
  • One human granulosa cell or fibroblast cell was
    injected into the perivitelline space of each of
    the enucleated bovine oocytes.

Injecting the cell
Cell placed subzonally
19
Materials and Methods
  • Treated oocytes were evaluated for evidence of
    cleavage and embryonic development daily.
  • Embryo quality was assessed using similar grading
    criteria to those employed in human IVF.

20
Results
1Number of embryos developed from the total
number of oocytes2Parthenogenetic
development3CEI Cloning Efficiency
IndexEmbryo success rate/ Parthenogenetic
success rate X100
21
Conclusions
  • Our results point out that SCNT as applied in the
    current study fusing human granulosa cells or
    fibroblast cells can be done.
  • The technique could be quite sensitive and
    predictive for similar SCNT attempts in humans
    for therapeutic or reproductive purposes.

22
First Human Cloned Embryo
  • This technology has enabled us to create the
    first human cloned embryo for reproductive
    purposes.

Zavos PM Human reproductive cloning the time is
near. Reproductive BioMedicine Online 6,
397398, 2003.
23
First Human Cloned Embryo
  • Nine human oocytes were enucleated.
  • Fused with whole human granulosa cells via
    electrical stimulation and activation.

24
First Human Cloned Embryo
  • The resulting cloned embryo reached the 8-10 cell
    stage and cryopreserved for future molecular
    analysis.

25
ANNOUNCEMENTThe First Fresh Cloned Embryo
Transfer in Human
  • We have produced and transferred the first fresh
    cloned embryo into the mother and we are awaiting
    for results
  • The mother is a 35 year-old woman
  • The embryo was properlyevaluated and
    transferredat a 4-cell stage
  • No pregnancy was established

26
Difficulties noted by Animal Cloners
  • Poor cloning response.
  • Poor implantation and pregnancy ratio.
  • Poor health of animals born.

27
Those difficulties are due to
  • Poorly designed experiments.(few animals used
    with no definite objective)
  • Poorly executed experiments.(hit and miss type
    of research)
  • Poorly approached experiments.(done under
    non-sterile and uncontrolled environments)
  • Poorly understood and interpreted.(when animals
    died, no clear view of their cause of death)

SOME DONE FOR FAME AND FORTUNE BY THE ANIMAL
CLONERS!
28
Ethics Morality Hypocrisy
LET US EXAMINE THE FACTS AS THEY APPEAR WITH THE
ANIMAL CLONERS! Hypocrisy in Action
29
Animal cloning is inefficient and is likely to
remain so for the foreseeable future (by Wilmut
Jaenisch, Time, 2001)
  • A number of studies have already demonstrated
    far higher rates of development, as measured in
    the proportion of live births to the number of
    embryos transferred, and in some cases matching
    or exceeding developmental rates seen in human
    IVF.

30
Nuclear-cytoplasmic interaction and development
of goat embryos reconstructed by nuclear
transplantation production of goats by serially
cloning embryos(Yong and Yuqiang Biol. Reprod.
58 266-269, 1998)
  • Embryos created 141
  • Live births 45
  • Success Rate () 32

Re-cloned goat embryos from a previous cloning
procedure (serially cloned embryos) Similar to
Current Human IVF Success Rates
31
Eight calves cloned from somatic cells of a
single adult (Kato et al, Science, 282 2095-8,
1998)
  • Embryos created 10
  • Live births 8
  • Success Rate () 80

32
Excerpts from the Congressional Hearings on Human
Cloning
  • Dolly is not normal. Dolly is overweight.
    (Jaenisch, 2001)

Dolly is obese!!!
  • Dolly may have subtle defects like in the brain.
    Dolly, I believe, is not normal. . we have no
    tests to check that. (Jaenisch, 2001)

Dolly has an IQ problem!!!
Under oath at the Congressional Hearing on Human
Cloning Research, March 28, 2001
33
Excerpts from President Bushs talk on Human
Cloning
  • Scientists wanting to do human cloning are going
    to create humans for spare parts and I am against
    that
  • (George W. Bush, President of the USA)

34
Excerpts from the Oxford Union Debate
  • You should be ashamed of yourself for wanting to
    clone a human being. You are going to create
    human monsters and you will fail. (Robert
    Winston, June 5, 2001)
  • Ironically, he said the exact same things about
    Robert Edwards 26 years ago about his efforts to
    create the first human being via IVF. Today,
    Winston embraced IVF and he is known in the UK as
    Mr. IVF.
  • That is hypocritical!!

35
Excerpts from the Oxford Union Debate
  • You should be ashamed of yourself for
    experimenting and killing human embryos. (Harry
    Griffin, Roslyn Institute, June 5, 2001)
  • Today, Mr. Griffin is given a license by the
    British Government and HFEA to kill human embryos
    and extract stem cells.
  • That is highly hypocritical!!!

36
Human-Bovine Hybrid Embryos Created
  • In order to avoid killing human embryos we have
    created the Human-Bovine hybrid embryo model to
    study various phenomena that needed to be
    evaluated during SCNT.
  • What has Mr. Griffin done to avoid killing human
    embryos?

37
Excerpts from the National Academy of Sciences
  • Animal cloning is inefficient and is likely to
    remain so for the foreseeable future
  • (Ian Wilmut, Roslyn Institute, August 2001)

Today the success is tremendous!! They continue
however to misrepresent the facts!
38
Excerpts from the National Academy of Sciences
  • Cloning will never be perfected and applied for
    human or animal purposes. It is impossible to
    reprogram 35,000 genes present in the human
    genome and yield a healthy human
    being. (Rudolph Jaenisch, MIT Professor, August
    2001)

Today we are cloning cattle commercially with 83
success rate astonishing!!
39
Another Experts Opinion on Human Cloning
.in monkeys the removal of the egg nucleus also
removes what Schatten called "molecular motors"
that are responsible for separating chromosomes
during cell division. He explained. "The cells
that result after those cell divisions all have
the wrong number of chromosomes."
We cannot do it in monkeys and therefore it
cannot be done in humans
40
Humans may be easier to clone than
animals!(August, 2001)
This is the first concrete genetic data showing
that the cloning process could be less
complicated in humans than in sheep Keith
Killian, Duke University Medical Center our
data show that you dont necessarily have these
problems (with the large offspring syndrome) in
humans. Randy Jirtle, Duke researcher
41
Response to Open Letter to British News Editors
by leading UK scientists
  • to reconsider the prominence given to repeated
    claims by certain scientists that they have
    cloned a human being, including those made by Dr.
    Panos Zavos last weekend. (21st January 2004)

We have NEVER claimed to have cloned a human
being!
42
Response to Open Letter to British News Editors
by leading UK scientists
  • none of those involved have produced a shred of
    evidence to substantiate their assertions .
    (21st January 2004)

We have published and continue to publish in
peer-reviewed scientific journals! It appears
that these leading scientists do not do their
homework nor READ and get informed before they
offer an opinion.
43
Publications
  • Zavos PM Human reproductive cloning the time is
    near. Reproductive BioMedicine Online 6, 397398,
    2003.
  • Illmensee K, Levanduski M, Zavos PM Development
    of an interspecies-specific bioassay using the
    bovine oocyte model to evaluate the potential of
    SCNT in humans. Journal of Assisted Reproduction
    and Genetics, 2004 (Accepted, in press, withdrawn
    due to leading UK scientists pressure).
  • Zavos PM, Illmensee K First Embryo Transfer of a
    Cloned Human Embryo. Middle East Fertility
    Society Journal (Submitted for publication).
  • Zavos PM, Illmensee K Human Reproductive
    Cloning The Post Mortem Effort. (Currently in
    preparation).

44
Scientific Presentations
  • The American Society for Reproductive Medicine,
    San Antonio, Texas, October 11-15, 2003.
  • The Austrian Society of Reproductive Medicine and
    Endocrinology, Bregenz, Austria, October 17-18,
    2003.
  • The Middle East Fertility Society, Beirut,
    December 10-13, 2003.
  • The 12th World Congress on Human Reproduction,
    Venice, Italy, March 14-16, 2005
  • The World DNA and Genome Day, Dalian, China,
    April 25-30, 2005.
  • The Indian International Conference on Update in
    Infertility, Bangalore, India, April 25-May 1,
    2005.
  • The American Society for Reproductive Medicine
    and the Canadian Fertility and Andrology Society,
    Montreal, Quebec, Canada, October 15-19, 2005
    (Submitted).

45
HFEA grants the first therapeutic cloning
license for research(11 August 2004)
Is it an act of God?
46
HFEA purposes
  • Increasing knowledge about the development of
    embryos
  • Increasing knowledge about serious disease
  • Enabling any such knowledge to be applied in
    developing treatments for serious disease

After destroying the embryo and its potential
for life
47
Therapeutic vs Reproductive Cloning in the UK
  • Human reproductive cloning is illegal in the UK.
    As a result of the Human Reproductive Cloning Act
    (2001) nobody in the UK is allowed to use cell
    nuclear replacement, or any other technique, to
    create a child.

But Therapeutic Cloning for Stem Cell Research is
allowed
48
Therapeutic Cloning Stem Cell Research vs
Reproductive Cloning
  • Stem cells from Living Human Embryos
  • Killing Dismembering Human Embryos
  • Grow stem cells in culture
  • Used in treatment of various diseases
  • Somatic (body) cells
  • Inject into enucleated oocytes
  • Induce embryo development
  • Used to create healthy babies for childless
    couples and treating infertility

49
The Future of Cloning
  • Further elucidation of the molecular mechanisms
    involved during the processes of embryogenesis
  • Careful tailoring of subsequently developed
    culture conditions and manipulation strategies
  • Appropriate screening methods

will eventually allow infertile couples to
safely have healthy, genetically related children
through Somatic Cell Nuclear Transfer (SCNT)
technology
50
This technology should be developed by scientists
and medical experts that
  • understand this type of work and the seriousness
    for its development
  • should be focused on carrying out this project,
    and
  • should work with leaders and governments at the
    International level, to ensure that this
    technology can be made safe and be disseminated
    properly

51
We intend to develop this technology by
  • Selecting appropriate cell lines for SCNT
  • Proper reprogramming the celllines in tissue
    culture prior to SCNT
  • Screening the cloned embryosprior to embryo
    transferinto recipients
  • Monitoring the ongoingpregnancies from the
    cloned embryos

52
Vilified, ridiculed, accused of perverting
nature.
But the cloning pioneers are in good company
Sunday Herald, Glasgow, Scotland, 10/21/01
53
The Future of Reproductive Cloning
Cloning, too, will probably come to be accepted
as a reproductive tool if it is carefully
controlled
Professor Robert Edwards, 2001
Sunday Herald, Glasgow, Scotland, 10/21/01
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