1. Cloning – In the Eyes of the Beholder Ida Chow, Ph.D. Society for Developmental Biology Bethesda, Maryland

2. Society for Developmental Biology The Society for Developmental Biology declares a voluntary five-year moratorium on cloning human beings, where ‘cloning human beings’ is defined as the duplication of an existing or previously existing human being by transferring the nucleus of a differentiated, somatic cell into an enucleated human oocyte, and implanting the resulting product for intrauterine gestation and subsequent birth. Sept. 1997. Extended Jan. 2003

3. Cloning Greek klon = twig (cuttings from one original plant) Molecular biology: copies of DNA (gene) fragments Cell cultures: cell multiplication by cell division (mitosis) Bacterial growth Plants budding Identical twins Identical genetic copies

4. Fundamental Question How does a single cell, the fertilized egg, transform into a complex organism, with many different types of cells, tissues and organs? - processes? - genes? - control / regulation?

6. Totipotency – Capable of giving origin to all the cell types in the organism, person, e.g., fertilized egg developing into a whole organism Gradual loss of the capability for differentiation with the process of development - Pluripotency, multipotency, e.g., superficial skin cell from deep layer skin cell

7. Definitions Somatic cells: body cells, full genetic complement, 2 sets of chromosomes = diploid, 46 in humans Germ cells: gametes, oocyte (unfertilized egg) and sperm, one half of genetic complement, 1 set of chromosomes = haploid, 23 in humans Fertilization = fusion of sperm and oocyte, full genetic complement, 1 set of chromosome from egg and 1 from sperm, diploid

8. Somatic Cell Nuclear Transfer Removal of the nucleus containing the genes of an unfertilized oocyte Fusion of an isolated body cell with the oocyte, or injection of the somatic cell nucleus into the oocyte Stimulation of the oocyte to begin the process of cell division and differentiation, producing the blastocyst (a hollowed ball with a mass of about 150 cells)

9. Fundamental Questions Reprogramming of gene expression: How to make the adult somatic cell nuclear DNA (genes) express the same way as in an embryo, that is, following the proper sequence. What molecules in the oocyte cytoplasm are capable of de-differentiating the adult somatic cell nucleus?

10. Somatic Cell Nuclear Transfer

12. Conditions Stroke Heart attacks Type I Diabetes Neural degenerative diseases Diseases of the blood Muscular dystrophies Damage to nerve cells Certain types of cancer Bone diseases

14. Differences Embryonic unlimited capability for cellular reproduction broader pluripotency more direct access to stem cells large number of available stem cells many identified inducing factors Adult limited capability for cellular reproduction limited pluripotency difficult access and identification of the stem cells limited number of available stem cells chromosomal defects

15. Reproductive Cloning Implantation of the blastocyst into a prepared womb of a female Further differentiation and growth of the cloned product Delivery of a full-term (or almost) cloned individual, with nuclear DNA identical to that of the donor of the body cell, and mitochondrial DNA identical to that of the oocyte; no genetic similarity to surrogate female if not oocyte donor

17. NO CLONING OF HUMAN BEINGS FOR REPRODUCTIVE PURPOSES

18. Figures in this presentation are from publications by the National Institutes of Health (http://www.nih.gov ): Stem Cells: a Primer (http://www.nih.gov/news/stemcell/primer.htm) Stem Cells: Scientific Progress and Future Research Directions (http://www.nih.gov/news/stemcell/scireport.htm)

19. Additional Resources Society for Developmental Biology (SDB): www.sdbonline.org, Focus on Stem Cells: http://sdb.bio.purdue.edu/publications/focus/index.htmlwww.sdbonline.org http://sdb.bio.purdue.edu/publications/focus/index.html National Institutes of Health (NIH): www.nih.gov, Stem Cell Information Index: http://www.nih.gov/news/stemcell/index.htmwww.nih.gov http://www.nih.gov/news/stemcell/index.htm National Bioethics Advisory Commission Reports (NBAC): http://www.georgetown.edu/research/nrcbl/nbac/pubs.html. Scrawl done and see the following reports: Ethical Issues in Human Stem Cell Research (Sept. 1999) and Cloning Human Beings (June 1997) http://www.georgetown.edu/research/nrcbl/nbac/pubs.html The President’s Council on Bioethics (PCBE): www.bioethics.gov, and see the Cloning Report: http://www.bioethics.gov/cloningreport/www.bioethics.gov http://www.bioethics.gov/cloningreport/ The National Academies: www.nas.edu. Scientific and Medical Aspects of Human Reproductive Cloning: http://www.nap.edu/catalog/10285.html?se_side, Stem Cells and the Future of Regenerative Medicine: http://www.nap.edu/catalog/10195.html?se_sidewww.nas.edu http://www.nap.edu/catalog/10285.html?se_side http://www.nap.edu/catalog/10195.html?se_side Federation of American Societies for Experimental Biology (FASEB): www.faseb.org, Cloning, Past, Present and the Exciting Future: http://www.faseb.org/opar/cloning/ www.faseb.org http://www.faseb.org/opar/cloning/