Therapeutic Cloning Research Paper

1. Snustad, Simmons . Principles of Genetics. 3. John Wiley & Sons Inc; 2003.

2. Illmensee K, Levanduski M, Zavos PM. Evaluation of the embryonic preimplantation potential of human adult somatic cells via an embryo interspecies bioassay using bovine oocytes. Fertil Steril. 2006 Apr;85(Suppl 1):1248–60.[PubMed]

3. Doss MX, Koehler CI, Gissel C, Hescheler J, Sachinidis A. Embryonic stem cells: a promising tool for cell replacement therapy. J Cell Mol Med. 2004 Oct–Dec;8(4):465–73. Review. [PubMed]

4. Arsanjani Mahnoush H. Negociating the UN Declaration on Human Cloning. The American Journal of International Law. 2006 Jan;100(1):164–179. accessed online from JSTOR archives.

5. Pattinson SD. University of Sheffield, Sheffield Institute of Biotechnological Law and Ethics (SIBLE). Some problems challenging the UK’s Human Fertilisation and Embryology Authority. Med Law. 2005 Jun;24(2):391–401.[PubMed]

6. Center for Genetics and Society. Canadian Parliament Approves the “Assisted Human Reproduction Act, “A Model of Responsible Policy.http://www.genetics-and-society.org/policies/other/canada.html.

7. Dennis C. Australia considers changing laws to allow therapeutic cloning. Nat Med. 2006 Feb;12(2):156.[PubMed]

8. Zavos PM, Illmensee K. Possible therapy of male infertility by reproductive cloning: one cloned human 4-cell embryo. Arch Androl. 2006 Jul–Aug;52(4):243–54.[PubMed]

9. Liang P, Jin LH, Liang T, Liu EZ, Zhao SG. Human neural stem cells promote corticospinal axons regeneration and synapse reformation in injured spinal cord of rats. Chin Med J (Engl) 2006 Aug 20;119(16):1331–8.[PubMed]

10. D’Amour KA, Bang AG, Eliazer S, Kelly OG, Agulnick AD, Smart NG, Moorman MA, Kroon E, Carpenter MK, Baetge EE. Production of pancreatic hormone-expressing endocrine cells from human embryonic stem cells. Nat Biotechnol. 2006 Oct[PubMed]

11. Barberi T, Klivenyi P, Calingasan NY, Lee H, Kawamata H, Loonam K, Perrier AL, Bruses J, Rubio ME, Topf N, Tabar V, Harrison NL, Beal MF, Moore MA, Studer L. Neural subtype specification of fertilization and nuclear transfer embryonic stem cells and application in parkinsonian mice. Nat Biotechnol. 2003 Oct;21(10):1200–7.[PubMed]

12. Sharpless NE, DePinho RA. Telomeres stem cells, senescence, and cancer. J Clin Invest. 2004;113:160–168.[PMC free article][PubMed]

13. Stampfer M, Garbe J, Levine G, Lichtsteiner S, Vasserot A, Yaswen P. Expression of the telomerase catalytic subunit, hTERT, induces resistance to transforming growth factor β growth inhibition in p16INK4A(−) human mammary epithelial cells. PNAS. 2001 Apr 10;98(8):4498–503.[PMC free article][PubMed]

14. Mitalipov SM, Wolf DP. Nuclear transfer in nonhuman primates. Methods Mol Biol. 2006;348:151–68.[PubMed]

15. Novak K. Therapeutic cloning gives silenced genes a second voice. Nat Med. 2004 Oct;10(10):100.[PubMed]

16. Hochedlinger K, Blelloch R, Brennan C, Yamada Y, Kim M, Chin L, Jaenisch R. Reprogramming of a melanoma genome by nuclear transplantation. Genes Dev. 2004 Aug 1;18(15):1875–85.[PMC free article][PubMed]

17. Li L, Connelly MC, Wetmore C, Curran T, Morgan JI. Mouse embryos cloned from brain tumors. Cancer Res. 2003 Jun 1;63(11):2733–6.[PubMed]

18. Fan J, Kodama E, Koh Y, Nakao M, Matsuoka M. Halogenated thymidine analogues restore the expression of silenced genes without demethylation. Cancer Research. 2005;65 (15):6927–6933.[PubMed]

19. Reik W, Romer I, Barton SC, Surani MA, Howlett SK, Klose J. Adult phenotype in the mouse can be affected by epigenetic events in the early embryo. Development. 1993 Nov;119(3):933–42.[PubMed]

20. Jaenisch R. Human cloning - the science and ethics of nuclear transplantation, Whitehead Institute for Biomedical Research. N Engl J Med. 2004 Dec 30;351(27):2787–91.[PubMed]

21. Blelloch R, Wang Z, Meissner A, Pollard S, Smith A, Jaenisch R. Reprogramming efficiency following somatic cell nuclear transfer is influenced by the differentiation and methylation state of the donor nucleus.1. Stem Cells. 2006 Sep;24(9):2007–13.[PMC free article][PubMed]

22. Liu W, Wang DR, Cao YL. TGF-beta: a fibrotic factor in wound scarring and a potential target for anti-scarring gene therapy. Curr Gene Ther. 2004 Mar;4(1):123–36. Review. [PubMed]

23. Huard J, Cao B, Qu-Petersen Z. Muscle-derived stem cells: potential for muscle regeneration. Birth Defects Res C Embryo Today. 2003 Aug;69(3):230–7. Review. [PubMed]

24. Rideout WM, 3rd, Hochedlinger K, Kyba M, Daley GQ, Jaenisch R. Correction of a genetic defect by nuclear transplantation and combined cell and gene therapy. Cell. 2002 Apr 5;109(1):17–27.[PubMed]

25. Hall VJ, Stojkovic P, Stojkovic M. Using therapeutic cloning to fight human disease: a conundrum or reality? Stem Cells. 2006 Jul;24(7):1628–37. Review. [PubMed]

26. Chinen J, Puck JM. Successes and risks of gene therapy in primary immunodeficiencies. J Allergy Clin Immunol. 2004 Apr;113(4):595–603. Review. [PubMed]

27. Acevedo B, Gomez-Palomares JL, Ricciarelli E, Hernández ER. Triggering ovulation with gonadotropin-releasing hormone agonists does not compromise embryo implantation rates. Fertility and Sterility. 2006 Dec;86(6):1682–7.[PubMed]

28. Mathur R, Kailasam C, Jenkins J. Review of the evidence base of strategies to prevent ovarian hyperstimulation syndrome. Human Fertility. 2007;10(2):75–85.[PubMed]

29. Okie S. Stem-cell research--signposts and roadblocks. N Engl J Med. 2005 Jul 7;353(1):1–5.[PubMed]

30. Hall VJ, Compton D, Stojkovic P, Nesbitt M, Herbert M, Murdoch A, Stojkovic M. Developmental competence of human in vitro aged oocytes as host cells for nuclear transfer. Human Reproduction. 2006 Sep 6;[PubMed]

31. Holden C. Stem cell research. Harvard cloners Get OK to proceed with caution. Science. 2006 Jun 16;312(5780):1584.[PubMed]

32. Nagy ZP, Chang CC. Artificial Gametes. Theriogenology. 2006 Oct 19;

33. Tesarik J, Nagy ZP, Sousa M, Mendoza C, Abdelmassih R. Fertilizable oocytes reconstructed from patient’s somatic cell nuclei and donor ooplasts. Reprod Biomed Online. 2001;2(3):160–164.[PubMed]

34. Dominko T, Mitalipova M, Haley B, Beyhan Z, Memili E, McKusick B, First NL. Bovine oocyte cytoplasm supports development of embryos produced by nuclear transfer of somatic cell nuclei from various mammalian species. Biol Reprod. 1999 Jun;60(6):1496–502.[PubMed]

35. Colman A, Kind A. Therapeutic cloning: concepts and practicalities. PPL Therapeutics, Trends Biotechnol. 2000 May;18(5):192–6.[PubMed]

36. Hall VJ, Stojkovic P, Stojkovic M. Using therapeutic cloning to fight human disease: a conundrum or reality? Stem Cells. 2006 Jul;24(7):1628–37. Review. [PubMed]

37. Sharov VG, Todor A, Khanal S, Imai M, Sabbah HN. Cyclosporine A attenuates mitochondrial permeability transition and improves mitochondrial respiratory function in cardiomyocytes isolated from dogs with heart failure. J Mol Cell Cardiol. 2006 Oct 26;[PMC free article][PubMed]

38. Park ES, Hwang WS, Jang G, Cho JK, Kang SK, Lee BC, Han JY, Lim JM. Incidence of apoptosis in clone embryos and improved development by the treatment of donor somatic cells with putative apoptosis inhibitors. Mol Reprod Dev. 2004 May;68(1):65–71.[PubMed]

39. Lanza RP, Cibelli JB, West MD. West Prospects for the use of nuclear transfer in human transplantation. Nature Biotechnology. 1999;17:1171–1174.[PubMed]

40. Yang XY, Li H, Ma QW, Yan JB, Zhao JG, Li HW, Shen HQ, Liu HF, Huang Y, Huang SZ, Zeng YT, Zeng F. Shanghai Institute of Medical Genetics. Improved efficiency of bovine cloning by autologous somatic cell nuclear transfer. Reproduction. 2006 Nov;132(5):733–739.[PubMed]

41. Lanza RP, Cibelli JB, West MD. Human therapeutic cloning. Nat Med. 1999 Sep;5(9):975–7.[PubMed]

42. Miyagawa S, Nakatsu S, Hazama K, Nakagawa T, Kondo A, Matsunami K, Yamamoto A, Yamada J, Miyazawa T, Shirakura R. A novel strategy for preventing PERV transmission to human cells by remodeling the viral envelope glycoprotein. Xenotransplantation. 2006 May;13(3):258–63.[PubMed]

43. Moalic Y, Blanchard Y, Felix H, Jestin A. Porcine endogenous retrovirus integration sites in the human genome: features in common with those of murine leukemia virus. Journal of Virology. 2006 Nov;80(22):10980–8.[PMC free article][PubMed]

44. Gruen L, Grabel L. Concise review: scientific and ethical roadblocks to human embryonic stem cell therapy. Stem Cells. 2006 Oct;24(10):2162–9.[PubMed]

45. Martin MJ, Muotri A, Gage F, Varki A. Glycobiology Research and Training Center and Department of Medicine. University of California. Human embryonic stem cells express an immunogenic nonhuman sialic acid. Nat Med. 2005 Feb;11(2):228–32.[PubMed]

46. Ludwig TE, Levenstein ME, Jones JM, Berggren WT, Mitchen ER, Frane JL, Crandall LJ, Daigh CA, Conard KR, Piekarczyk MS, Llanas RA, Thomson JA. WiCell Research Institute. Derivation of human embryonic stem cells in defined conditions. Nat Biotechnol. 2006 Feb;24(2):185–7.[PubMed]

47. Fujikawa T, Oh SH, Pi L, Hatch HM, Shupe T, Petersen BE. Teratoma formation leads to failure of treatment for type I diabetes using embryonic stem cell-derived insulin -producing cells. American Journal of Pathology. 2005 Jun;166(6):1781–91.[PMC free article][PubMed]

48. Mimeault M, Batra SK. Recent Advances on the Significance of Stem Cells in Tissue Regeneration and Cancer Therapies. Stem Cells. 2006;24(11):2319–2345.[PubMed]

49. Ostrer H, Wilson DI, Hanley NA. Human embryo and early fetus research. Clin Genet. 2006 Aug;70(2):98–107.[PubMed]

50. Landry DW, Zucker HA. Embryonic death and the creation of human embryonic stem cells. J Clin Invest. 2004 Nov;114(9):1184–6. Review. [PMC free article][PubMed]

51. The President’s Council on Bioethics. Washington, D.C: Jul, 2002. Human Cloning and Human Dignity: An Ethical Inquiry. position number one of the section entitled “the Moral Case for Cloning-for-Biomedical-Research.” http://www.bioethics.gov/reports/cloningreport/fullreport.html.

52. Alberio R, Campbell KH, Johnson AD. Reprogramming somatic cells into stem cells. Reproduction. 2006 Nov;132(5):709–20.[PubMed]

53. Wakayama T. On the road to therapeutic cloning. Nat Biotechnol. 2004 Apr;22(4):399–400.[PubMed]

54. Roberts PC, Lowe C. Where have all the conceptions gone? Lancet. 1975:498–499.

55. Singer P, Dawson Karen. IVF Technology and the Argument from Potential. Philosophy and Public Affairs. 1988;17(2):89–90.[PubMed]

56. McMahan J. The Ethics of Killing - Problems at the Margins of Life. Oxford University Press; 2002. p. 308.

57. Lanza RP, Caplan AL, Silver LM, Cibelli JB, West MD, Green RM. The ethical validity of using nuclear transfer in human transplantation. JAMA. 2000 Dec 27;284(24):3175–9. Ethics Institute, Dartmouth College, Hanover. [PubMed]

58. Meissner A, Jaenisch R. Generation of nuclear transfer-derived pluripotent ES cells from cloned Cdx2-deficient blastocysts. Nature. 2006 Jan 12;439(7073):212–5.[PubMed]

Scientists at Harvard and the affiliated Children’s Hospital Boston announced on June 6 that they had begun experiments with somatic cell nuclear transfer (SCNT). By combining donated human eggs with skin cells from patients who have sickle-cell anemia or diabetes, the scientists will attempt to clone diseased cells and then to derive stem-cell lines from those cloned cells. If successful, the efforts will result in stem-cell lines that carry specific diseases, and thus rapidly accelerate research into causes and cures.

Because of the highly charged ethical and political debate over the work, scientists George Q. Daley ’82, M.D., Kevin Eggan, and Douglas Melton received permission to pursue the research only “after more than two yearsof intensive review by eight different institutional review boards and stem-cell oversight committees at five different institutions,” said University provost Steven E. Hyman. “This highly unusual collaborative effort of the Harvard Stem Cell Institute,” he noted, “involves researcher sat Columbia University Medical Center’s Naomi Berrie Diabetes Center, Boston IVF [a center for assisted reproduction], the Center for Reproductive Medicine at Brigham and Women’s Hospital, and the New York Stem Cell Foundation.” Because of the Bush administration ban on federal funding of stem-cell lines created after August 9, 2001, “there is no taxpayer money being used for any embryonic stem-cell work,” Hyman added. “This is all [made possible by] private donation.”

For his research into diseases of the blood, Daley, of Children’s Hospital Boston (and an associate professor of medicine at Harvard Medical School), will use frozen, failed-to-fertilize eggs donated by couples who have ended their fertility treatments. Faculty of Arts and Sciences professors Eggan and Melton, tackling diabetes, will instead seek donations of fresh eggs. “We don’t know which strategy will work in the long run,” Daley said. For example, the researchers anticipate, but don’t know, that fresh eggs may be harder to come by; previously frozen eggs, on the other hand—those that failed to fertilize during assisted reproductive efforts—might not function. By taking “separate but complementary approaches and by working together and communicating,” Daley said, “our hope is that we will actually answer these questions.”

Having performed somatic cell nuclear transfer successfully with mous embryonic stem cells (above), Harvard researches are now starting to work with human cells.
To ensure that egg donors' consent is fully informed, the Harvard Stem Cell Institute created a handbook for donors. Freshly donated eggs, rather than frozen ones, may be necessary for successful nuclear transfer.

Although no one has yet published a credible paper reporting the derivation of a human embryonic stem-cell line from a blastocyst created by nuclear transfer, the technological hurdles appear surmountable. A group in the United Kingdom has reported success with SCNT using fresh eggs, but made no attempt to derive a stem-cell line from the resulting cell mass, which was destroyed instead as part of a definitive proof that the nuclear transfer had succeeded. Melton, in an earlier collaboration with Boston IVF and the laboratory of Andrew McMahon, Baird professor of science, has created more than 30 human em- bryonic stem-cell lines for free distribution to scientists around the world. Separately at least, then, the two halves of the problem Harvard scientists now propose to tackle have been solved (see box). Successfully bringing them together would create a powerful new tool against diseases. Because stem cells are self-renewing once isolated in a petri dish, scientists would be able to set out thousands of samples of diseased cells and then test different drug compounds on each sample to see if one cured the disease.

Stem-cell scientists (from top) Douglas Melton, George Q. Daley, and Kevin Eggan.

Another advantage of such stem-cell lines is the opportunity they present for “homologous recombination”: gene therapy in a petri dish. Daley has already used this technique to successfully restore partial immune function in a mouse with severe blood disease. In that experiment, he cloned a cell from the mouse’s tail, derived a stem-cell line, repaired the defective DNA in a petri dish, verified that it had been fixed, and then put corrected cells back into the mouse, where they reconstituted its blood and immune system. As a practicing physician, he hopes one day to use this therapeutic cloning technique to cure his patients.

The diseases the scientists hope to study, such as diabetes and Parkinson’s, are frequently the result of complex interactions among multiple genes and the environment. That means “it is impossible in the laboratory to use established molecular techniquesto make cell lines or animal models for [study],” said Eggan. “In fact, it is difficult even to identify [such] patients using genetic methods until they step into the clinic with the disease. This is one of the only methods we can imagine using for making cell-based models of these diseases.”

Opponents object to such work on ethical grounds: they believe it involves the destruction of an embryo that could become a person. That is scientifically debatable, because a sperm-fertilized egg starts dividing immediately and thus has the capacity for self-directed growth, whereas cloned cells are coaxed into cell division by scientists using chemical or electrical stimuli.

Of much greater ethical concern to the oversight committees that reviewed the Harvard team’s proposal was egg donation (which clouded the now discredited therapeutic cloning work of South Korean scientist Hwang Woo Suk). Egg donation involves a series of carefully timed injections with three different hormones, sexual abstinence for a month, frequent visits to a nurse, and a surgical procedure under anesthesia to retrieve the eggs. To avoid even the appearance of improper inducement, egg donors will not be compensated, except for their expenses. And to ensure that their consent is fully informed, the Harvard Stem Cell Institute has created the Research Egg Donor Information Book to explain the research and the medical risks. Who might be willing to donate? “At this point,” said Eggan, “we might guess that women [whose family members] are afflicted with diseases that we are interested in studying might step forward to participate. That is our best hope.”

Therapeutic cloning is a two-step process. Scientists must first transfer the nucleus of a skin (or other somatic) cell that they want to clone into an enucleated egg (one that has had its nucleus—hence all its DNA—removed). This is extremely difficult to execute successfully: in effect, it is performing surgery on a single cell. The second part of the process, derivation of a stem-cell line from the new cell created by the nuclear transfer, is also challenging. Scientists coax the cloned cell to divide for several days until they have a mass of four to 50 undifferentiated cells. Stem cells must be harvested from this cluster, called a blastocyst, at just the right moment; researchers have about a 24-hour window during which this fleeting cell-type can be successfully transferred, to be grown indefinitely in culture on a bed of feeder cells.

0 Replies to “Therapeutic Cloning Research Paper”

Lascia un Commento

L'indirizzo email non verrà pubblicato. I campi obbligatori sono contrassegnati *