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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

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slide1

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

introduction
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.
slide3

“Public hostility to human reproductive cloning may be based on an illogical transient fear of a new technology”

The British Medical Association

slide4

Infertility is a disease

Today, infertility is a disease that reaches epidemic proportions throughout the developing World

slide5

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

slide6

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

slide7

Sperm and Oocyte Donation

  • No mature sperm present
  • No oocyte (egg) production

Offspring not genetically or biologically related

slide8

Quotes from childless patients

  • “..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)
slide9

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.

slide10

Sexual and Asexual Reproduction

No difference in the type of oocyte (egg) used

slide11

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”

slide12

Sexual and Asexual Reproduction

No difference in the cell division stages after fertilization

slide13

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.
slide14

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
slide15

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.
slide16

Materials and Methods

  • Bovine oocytes were enucleated by aspiration of the first polar body and the metaphase plate.
slide17

Materials and Methods

  • Human granulosa cells and fibroblast cells were aspirated into a micropipette.
slide18

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

slide19

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.
slide20

Results

1Number of embryos developed from the total number of oocytes;2Parthenogenetic development;3CEI: Cloning Efficiency Index=Embryo success rate/ Parthenogenetic

success rate X100

slide21

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.
slide22

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, 397–398, 2003.

slide23

First Human Cloned Embryo

  • Nine human oocytes were enucleated.
  • Fused with whole human granulosa cells via electrical stimulation and activation.
slide24

First Human Cloned Embryo

  • The resulting cloned embryo reached the 8-10 cell stage and cryopreserved for future molecular analysis.
slide25

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
slide26

Difficulties noted by “Animal Cloners”

  • Poor cloning response.
  • Poor implantation and pregnancy ratio.
  • Poor health of animals born.
slide27

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!

slide28

Ethics Morality & Hypocrisy

LET US EXAMINE THE FACTS AS THEY APPEAR WITH THE ANIMAL CLONERS!

Hypocrisy in Action

slide29

“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.

slide30

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

slide31

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%

slide32

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

slide33

Excerpts from President Bush’s 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)
slide34

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!!
slide35

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!!!
slide36

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?
slide37

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!

slide38

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!!

slide39

Another “Expert’s” 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”

slide40

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 don’t necessarily have these problems (with the large offspring syndrome) in humans.” Randy Jirtle, Duke researcher

slide41

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!

slide42

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.

slide43

Publications

  • 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).
slide44

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).
slide45

“HFEA grants the first therapeutic cloning license for research”(11 August 2004)

Is it an act of God?

slide46

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

slide47

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

slide48

Therapeutic Cloning & Stem Cell Research vs Reproductive Cloning

  • Somatic (body) cells

Inject into enucleated oocytes

  • Induce embryo development
  • Used to create healthy babies for childless couples and treating infertility
  • Stem cells from Living Human Embryos
  • Killing & Dismembering Human Embryos
  • Grow stem cells in culture
  • Used in treatment of various diseases
slide49

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

slide50

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
slide51

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
slide52

Vilified, ridiculed, accused of perverting nature….

But the cloning pioneers are in good company

Sunday Herald, Glasgow, Scotland, 10/21/01

slide53

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