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Professor Dr. Panos Zavos, Ed.S., Ph.D.Professor Dr. Panos Zavos, Ed.S., Ph.D.
TO CLONE OR NOT TO CLONE?
WHOSE CHOICE IS IT ANYWAY?
TO CLONE OR NOT TO CLONE?
WHOSE CHOICE IS IT ANYWAY?
Professor Emeritus of Reproductive Physiology/Andrology, University of Kentucky Director, Andrology Institute of AmericaAssociate Director, Kentucky Center for Reproductive Medicine & IVF, Lexington, KY, USAExecutive Director, The Zavos Organization
Professor Emeritus of Reproductive Physiology/Andrology, University of Kentucky Director, Andrology Institute of AmericaAssociate Director, Kentucky Center for Reproductive Medicine & IVF, Lexington, KY, USAExecutive Director, The Zavos Organization
IntroductionIntroduction
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.
The British Medical AssociationThe British Medical Association
“Public hostility to human reproductive cloning may be
based on an illogical transient fear of a new
technology”
“Public hostility to human reproductive cloning may be
based on an illogical transient fear of a new
technology”
Today, infertility is a disease that reaches epidemic proportions throughout the developing World
Infertility is a diseaseInfertility is a disease
Low sperm count and/or motility
Variety of female factors
Success rate ~ 33% live birth/transfer
Did not overcome severe asthenospermia
In Vitro Fertilization (IVF)In Vitro Fertilization (IVF)
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
Intra Cytoplasmic Sperm Injection (ICSI)
Intra Cytoplasmic Sperm Injection (ICSI)
No mature sperm present No oocyte (egg) production
Offspring not genetically or biologically related
Sperm and Oocyte DonationSperm and Oocyte Donation
“..we want a child (yesterday, if possible) and a healthy child.”
“..do not want to have another person’s spermor eggs.”
“..what other options do we have?”
“…we want to have a biological child of our own.”
(Received via e-mail from patients)
Quotes from childless patients
Quotes from childless patients
1. First, the nucleus of a donor egg is removed.
2. Then a whole somatic cell or the nucleus of a whole somatic cell from a patient is inserted.
3. The result is an egg with the patient's genetic material.
4. 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.
How Therapeutic Cloning Works
How Therapeutic Cloning Works
No difference in the type of oocyte (egg) used
Sexual and Asexual Reproduction
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”
Sexual and Asexual Reproduction
Sexual and Asexual Reproduction
No difference in the cell division stages after
fertilization
Sexual and Asexual Reproduction
Sexual and Asexual Reproduction
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.
Cloning in AnimalsCloning in Animals
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
Our StudiesOur Studies
Bovine oocytes were randomly treated either for induction of parthenogenesis or for enucleation and SCNT using either human granulosa cells or human fibroblast cells.
Materials and MethodsMaterials and Methods
Bovine oocytes were enucleated by aspiration of the first polar body and the metaphase plate.
Materials and MethodsMaterials and Methods
Human granulosa cells and fibroblast cells were aspirated into a micropipette.
Materials and MethodsMaterials 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
Materials and MethodsMaterials 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.
Materials and MethodsMaterials and Methods
Experiment 1 Experiment 2
Granulosa cells
Controls 2 Fibroblast cells
Controls
2
Success rate1
31.2% (15/48)
46.8% (36/77)
24.2 % (15/62)
56.8% (21/37)
CEI 3 66% 42%1Number of embryos developed from the total number of oocytes;2Parthenogenetic development;3CEI: Cloning Efficiency Index=Embryo success rate/ Parthenogenetic success rate X100
ResultsResults
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.
ConclusionsConclusions
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, 397–398, 2003.
First Human Cloned EmbryoFirst Human Cloned Embryo
Nine human oocytes were enucleated.
Fused with whole human granulosa cells via electrical stimulation and activation.
First Human Cloned EmbryoFirst Human Cloned Embryo
The resulting cloned embryo reached the 8-10 cell stage and cryopreserved for future molecular analysis.
First Human Cloned EmbryoFirst Human Cloned Embryo
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
ANNOUNCEMENTThe First Fresh Cloned
Embryo Transfer in Human
ANNOUNCEMENTThe First Fresh Cloned
Embryo Transfer in Human
Poor cloning response. Poor implantation and pregnancy ratio. Poor health of animals born.
Difficulties noted by “Animal Cloners”
Difficulties noted by “Animal Cloners”
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)
Those difficulties are due to:Those difficulties are due to:
SOME DONE FOR FAME AND FORTUNE BY THE ANIMAL CLONERS!
Ethics Morality & HypocrisyEthics Morality & Hypocrisy
LET US EXAMINE THE FACTS AS THEY APPEAR
WITH THE ANIMAL CLONERS!
Hypocrisy in Action
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.
“Animal cloning is inefficient and is likely to remain so for
the foreseeable future” (by Wilmut & Jaenisch, Time, 2001)
“Animal cloning is inefficient and is likely to remain so for
the foreseeable future” (by Wilmut & Jaenisch, Time, 2001)
Embryos created* 141
Live births 45
Success Rate (%) 32%**
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)
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)
*Re-cloned goat embryos from a previous cloning procedure (serially cloned embryos)
**Similar to Current Human IVF Success Rates
Embryos created 10
Live births 8
Success Rate (%) 80%
Eight calves cloned from somatic cells of a single adult
(Kato et al, Science, 282: 2095-8, 1998)
Eight calves cloned from somatic cells of a single adult
(Kato et al, Science, 282: 2095-8, 1998)
• “Dolly is not normal. Dolly is overweight.” (Jaenisch, 2001)
*Under oath at the Congressional Hearing on Human Cloning Research, March 28, 2001
Dolly is obese!!!
Dolly has an IQ problem!!!
• “Dolly may have subtle defects like in the brain. Dolly, I believe, is not normal. …. we have no tests to check that.” (Jaenisch, 2001)
Excerpts from the Congressional Hearings on
Human Cloning*
Excerpts from the Congressional Hearings 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)
Excerpts from President Bush’s talk on Human
Cloning
Excerpts from President Bush’s talk on Human
Cloning
• “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!!
Excerpts from the Oxford Union Debate
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!!!
Excerpts from the Oxford Union Debate
Excerpts from the Oxford Union Debate
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?
Human-Bovine Hybrid Embryos Created
Human-Bovine Hybrid Embryos Created
• “Animal cloning is inefficient and is likely to remain so for the foreseeable future”
(Ian Wilmut, Roslyn Institute, August 2001)
Excerpts from the National Academy of Sciences
Excerpts from the National Academy of Sciences
Today the success is tremendous!!
They continue however to misrepresent the facts!
• “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)
Excerpts from the National Academy of Sciences
Excerpts from the National Academy of Sciences
Today we are cloning cattle commercially with 83% success rate;
astonishing!!
“….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."
Another “Expert’s” Opinion on Human Cloning
Another “Expert’s” Opinion on Human Cloning
“We cannot do it in monkeys and therefore it cannot be done in
humans”
“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 don’t necessarily have these problems (with the large offspring syndrome) in humans.” Randy Jirtle, Duke researcher
Humans may be easier to clone than animals!
(August, 2001)
Humans may be easier to clone than animals!
(August, 2001)
“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 We have NEVER claimed to have cloned a human being!cloned a human being!
Response to Open Letter to British News Editors by “leading UK scientists”
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 We have published and continue to publish in peer-reviewed scientific journals!peer-reviewed scientific journals!
It appears that these “leading scientists” do It appears that these “leading scientists” do not do their homework nor READ and get not do their homework nor READ and get
informed before they offer an opinion.informed before they offer an opinion.
We have published and continue to publish in We have published and continue to publish in peer-reviewed scientific journals!peer-reviewed scientific journals!
It appears that these “leading scientists” do It appears that these “leading scientists” do not do their homework nor READ and get not do their homework nor READ and get
informed before they offer an opinion.informed before they offer an opinion.
Response to Open Letter to British News Editors by “leading UK scientists”
Response to Open Letter to British News Editors by “leading UK scientists”
Zavos PM: Human reproductive cloning: the time is near. Reproductive BioMedicine Online 6, 397–398, 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).
PublicationsPublications
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).
Scientific Presentations
Scientific Presentations
Is it an act of God?Is it an act of God?
“HFEA grants the first therapeutic cloning license
for research”(11 August 2004)
“HFEA grants the first therapeutic cloning license
for research”(11 August 2004)
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 After destroying the embryo and its potential for lifeits potential for life
HFEA purposesHFEA purposes
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 But Therapeutic Cloning for Stem Cell Research is allowedResearch is allowed
Therapeutic vs Reproductive Cloning in
the UK
Therapeutic vs Reproductive Cloning in
the UK
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
Therapeutic Cloning & Stem Cell Research vs Reproductive
Cloning
Therapeutic Cloning & Stem Cell Research vs Reproductive
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
The Future of CloningThe Future of Cloning
will eventually allow infertile couples to safely have healthy, genetically
related children through Somatic Cell Nuclear Transfer (SCNT) technology
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
This technology should be developed by scientists and
medical experts that:
This technology should be developed by scientists and
medical experts that:
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
We intend to develop this technology by:
We intend to develop this technology by:
Sunday Herald, Glasgow, Scotland, 10/21/01
Vilified, ridiculed, accused of perverting nature….
Vilified, ridiculed, accused of perverting nature….
But the cloning pioneers are in good company
But the cloning pioneers are in good company
“Cloning, too, will probably come to be accepted as a reproductive tool if it is carefully
controlled” Professor Robert Edwards,
2001Sunday Herald, Glasgow, Scotland, 10/21/01
The Future of Reproductive Cloning
The Future of Reproductive Cloning
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