DAVID WELLS

Nomination Statement for McMeekan Award :
Research in the last 5 years

David is currently a highly valued Scientist in the Reproductive Technologies Group at AgResearch, Ruakura Agricultural Research Centre in Hamilton. He is leader of the Foundation of Research Science and Technology (FRST) funded NERF objective on cloning of livestock from adult somatic cells. The objective has achieved outstanding results over the last 5 years and for this reason, David is being nominated for the McMeekan Award.

Award Achievements are:

· Established lines of differentiated somatic cells from sheep embryos, cattle late-stage embryos (34 days after conception) and adult cattle.

· Demonstration that these cell lines are totipotent using the technique of nuclear transfer. This involves firstly synchronising the cells into particular stages of the cell cycle, then transferring individual cells into the peri-vitelline space of eggs which have had their own chromosomes removed, electrically fusing the egg and cell cytoplasm and activating the embryo (either electrically or chemically) so that the embryo develops to the blastocyst stage in vitro. Blastocysts of suitable quality are then transferred to recipient animals which carry the embryos to term. The resulting offspring are clones of the somatic cell line as the genetic contribution is entirely from the cell fused to the egg cytoplasm.

· Investigating factors affecting the efficiency of the nuclear transfer procedures. This has resulted in technology which results in an approximately two to five-fold increase in efficiency of producing cattle clones compared to the best overseas groups. This is a very significant achievement, and combining his technology with that of the dairy cattle transgenics programme puts New Zealand at the forefront of international biotechnology research. This has been recognised by the overseas biotechnology companies, Pharmaceutical Proteins Ltd, UK, and Geron (Clone International), who have established joint ventures in New Zealand with AgResearch in pharmaceutical production from transgenic dairy cattle and animal cloning, respectively.

· David also filed a patent for novel nuclear transfer technology (NZ patent no. 337792).

· Transferred cloned embryos have resulted in the birth of the following animals:

o Lambs from both male and female cultured embryonic cell lines. The original lambs produced were the second report in the world of animals born following nuclear transfer of somatic cells. The lambs have now reached puberty and have been shown to be fertile. Furthermore, mating of cloned rams and cloned ewes has demonstrated normal sexual reproduction.

o Calves from cell lines established from a late stage embryo. These calves were the first born in the Southern Hemisphere from somatic cell cloning and the third report in the world of such cloning.

o Calves from somatic cells derived from adult cattle. These calves are the second report in the world (the first report occurring only 6 weeks previously) of animals produced from adult cells. A unique component of the cloning work was that the calves were clones of an endangered Enderby Island cow and therefore the results demonstrated the potential of the technology for rescuing endangered species.

o 19 calves from somatic cells derived from an adult high breeding worth Friesian dairy cow. These calves represented the first, largest single group of cloned offspring born from a living animal. These animals (as well as subsequent offspring) are being monitored for production characteristics as they age.

o In total over 70 cloned offspring have been produced following somatic cell nuclear transfer with a variety of different cell lines.

· The above achievements are outstanding, especially when it is realised that a small team of people (originally David plus one Research Associate. Now Dave plus 7 staff) conducted all the laboratory work. This indicates the enthusiasm, dedication and skill of David in his chosen work area.

· In his role as head of nuclear transfer research, David has a number of internal AgResearch collaborations. These include a joint project with Dr Phil L'Huillier and Goetz Laible, leaders of the dairy cattle transgenics programme, which has led to a number of births of cloned/transgenic cattle produced from adult cells transformed with bovine genes of commercial importance. Calves born include 11 with overexpression of b and k casein and four transgenic with the human Myelin Basic Protein gene. Other collaborations are with: Dr William Vivanco, Manager of arTech and leader of the Meat New Zealand funded programme on embryonic cloning; Dr Rita Lee, investigating the molecular biology of placental formation and imprinting; Dr Goetz Laible, investigating gene methylation patterning and Dr Chris Morris, investigating the population genetics of puberty and other performance traits. These collaborations have led to pregnancies of cloned cattle and sheep. A further recent collaboration is with Wayne Hein, Wallaceville, to produce clones of high and low parasite resistant lines of sheep for Imprivo, a company studying immunology of disease resistance.

· David also has important international collaborative links with overseas science groups and, as already mentioned, with both PPL Ltd and Geron BioMed, two leading biotechnology companies. He has assisted the Business Group in successfully negotiating exclusive licenses with these companies in Australasia, thereby ensuring the competitiveness of New Zealand's fledging biotechnology industries.

· Further collaborations are with Profs Nieman (Germany, gene expression in clones); Piedrahita (USA, transgenics); and Bren (Austria, nuclear transfer).

· David's achievements in somatic cell cloning have been recognised by the international science community through invitations to present results at the following conferences:

o 13th Brazilian Congress on Animal Reproduction, Belo Horizonte, Brazil, 26-30 July 1999.

o Genetically engineering and cloning animals, science, society and industry, Utah, USA. June 17-20, 2000.

o The "Cloning Symposium: Reprogramming Cell Fate - Transgenesis and Cloning", Monash University, Melbourne, 14-16 April, 1998.

o "New Zealand Institute of Medical Laboratory Science" meeting, Palmerston North, September 1-4, 1998.

o In addition, numerous press/media conferences have been held to announce the results and considerable correspondence has been entered into to describe the technology to members of the public. This includes discussions with teachers, students, governmental advisory groups, industry groups, general public and assistance with high school textbooks.

o Numerous presentations describing the technology have been made at national and international conferences.

o Other invitations have been to act as a consultant to commercial cloning organisations.

o David has published/submitted 51 papers, abstracts and general articles (see attached bibliography) over the last 5 years.

The above shows that David Wells, with a small team of staff, has developed world-leading nuclear transfer technology which underpins New Zealand's entry into the high value transgenic pharmaceutical/nutraceutical and animal cloning industries. This success, in a very competitive international field, mirrors David's skills at research plus his dedication and enthusiasm for his chosen research field. Seldom does New Zealand produce such an internationally recognised scientist, and for this reason David is an extremely worthy recipient of the McMeekan Award.

Robin Tervit …………………………….

John Smith ………………………………

17 December 2001.

PUBLICATIONS

Wells, D.N.; Thompson, J.G.E.; Tervit, H.R.; James, R.W.; Udy, G.B. 1990. Experiences in the application of embryo bisection in sheep MOET programmes. The proceedings of the New Zealand Society of Animal Production 50: 431-435.

Wells, D.N.; Kimber, S.J.; Wilmut, I. 1990. Cell surface carbohydrate antigens expressed during ovine preimplantation embryogenesis. Journal of Reproduction and Fertility, Abstract Series 5: 23.

Wells, D.N.; McWhir, J.; Hooper, M.L.; Wilmut, I. 1991. Factors influencing the isolation of murine embryonic stem cells. Theriogenology 35: 293.

Wells, D.N.; Wilmut, I.; McWhir, J.; Hooper, M.L. 1991. Is an epigenetic modification required for the isolation into culture, of embryonic stem cells? Proceedings of the EMBL Mouse Molecular Genetics Conference: Abstract 222.

Wells, D.N.; Wilmut, I.; McWhir, J.; Hooper, M.L. 1991. Is an epigenetic modification required for the isolation of murine embryonic stem cells into culture? Proceedings of the European Embryo Transfer Society 7: 218.

Wells, D. 1993. The in vitro isolation of murine embryonic stem cells. In: Methods in Molecular Biology, Volume 18: Transgenesis Techniques: Principles and Protocols, Murphy, D. and Carter, D.A. (eds), Humana Press, Totawa, NJ, pages 183-216.

Wells, D. 1993. Production of chimaeras derived from murine embryonic stem cells. In: Methods in Molecular Biology, Volume 18: Transgenesis Techniques: Principles and Protocols, Murphy, D. and Carter, D.A. (eds), Humana Press, Totawa, NJ, pages 217-237.

Parkes, B.D.; Udy, G.B.; Wells, D.N. 1994. Embryonic stem (ES) cell line has a major effect on targeting efficiency of the hypoxanthine-guanine phosphoribosyl transferase (HPRT) gene. Queenstown Molecular Biology Meeting 4: 138.

Wells, D.; Driessen, C.A.; Udy, G.B.; Tervit, H.R. 1995. Putative ovine embryonic stem cells grow in media containing different levels of energy substrates. Theriogenology 43: 349.

Udy, G.B.; Wheeler, T.T.; Wells, D.N.; Parkes, B.D. 1995. Variable ES cell phenotypes and gene targeting frequencies: a view of the early stem cell continuum? Queenstown Molecular Biology Meeting 5: 112.

Udy, G.B.; Wells, D.N. 1996. Low oxygen atmosphere initially increases the survival and multiplication of putative goat embryonic stem cells. Theriogenology 45: 237.

Misica, P.M.; Wells, D.N.; Tervit, H.R.; Thompson, J.G. 1996. Oxygen uptake by component tissues of blastocyst stage ovine embryos and putative embryonic stem cells: a preliminary study. International Congress on Animal Reproduction: Techniques for Gamete Manipulation and Storage 13: 23.

Udy, G.B.; Parkes, B.D.; Wells, D.N. 1997. ES cell cycle rates affect gene targeting frequencies. Experimental Cell Research 231: 296-301.

Wells, D.N.; Misica, P.M.; Day, A.M.; Tervit, H.R. 1997. Production of cloned lambs from an established embryonic cell line: a comparison between in vivo- and in vitro-matured cytoplasts. Biology of Reproduction 57: 385-393.

Wells, D.N.; Misica, P.M.; Day, A.M.; Tervit, H.R. 1997. Generation of cloned lambs from an established embryonic fibroblast cell line. Proceedings of Transgenic Animals in Agriculture Conference, Abstract 18.

Misica, P.M.; Peterson, A.J.; Day, A.M.; Wells, D.N. 1998. Co-transfer of trophoblastic vesicles with cloned sheep embryos is unable to improve survival to term. Theriogenology 49: 247.

Wells, D.N.; Misica, P.M.; McMillan, W.H.; Tervit, H.R. 1998. Production of cloned bovine fetuses following nuclear transfer with cells from a fetal fibroblast cell line. Theriogenology 49: 330.

Wells, D.N.; Misica, P.M.; Day, A.M.; Peterson, A.J.; Tervit, H.R. 1998. Cloning sheep from cultured embryonic cell lines. The Cloning Symposium: Reprogramming Cell Fate - Transgenesis and Cloning, Monash Medical Centre, Melbourne, April 15-16. Abstract.

Wells, D.N.; Misica, P.M.; Tervit, H.R. 1998. Cloning livestock animals from cultured cells using nuclear transfer. Genetically Engineering and Cloning Animals: Science, Society and Industry, Park City, Utah, June 21-23. Conference paper #2.

Wells, D.N.; Misica, P.M.; Tervit, H.R. 1998. Future opportunities in livestock production and biomedicine from advances in animal cloning. Proceedings of the New Zealand Society of Animal Production 58: 32-35.

Wells, D.N. 1998. Agricultural and biomedical applications from recent advances in animal cloning technology. Proceedings of the New Zealand Institute of Medical Laboratory Science, Palmerston North, September 1-4, Abstract: 22.

Tervit, H.R.; Vivanco, H.W.; Wells, D.N. 1998. Embryo technology for the dairy industry. New Zealand Veterinary Association Conference 184: 235-244.

Wells, D.N. 1998. Future opportunities in livestock production and biomedicine from advances in animal cloning. The Chamber Voice; Waikato Chamber of Commerce and Industry. Issue 11: 1-3.

Wells, D.N.; Misica, P.M.; Forsyth, J.T.; Berg, M.C.; Lange, J.M.; Tervit, H.R.; Vivanco, W.H. 1999. The use of adult somatic cell nuclear transfer to preserve the last surviving cow of the Enderby Island cattle breed. Theriogenology 51: 217.

Wells, D.N.; Misica, P.M.; Tervit, H.R. 1999. Production of cloned calves following nuclear transfer with cultured adult mural granulosa cells. Biology of Reproduction 60: 996-1005.

Wells, D.N. 1999. Cloning and transgenic technology to produce genetically enhanced dairy cattle in the future. Dairying Research Corporation Farmers' Field Day, March 11 1999, pages 10-12.

Wells, D. 1999. Cloned and transgenic dairy cattle. Large Herds Dairy Conference, Whakatane, March 16, Abstract.

Wells, D.N.; Misica, P.M.; Tervit, H.R.; Vivanco, W.H. 1999. Adult somatic cell nuclear transfer is used to preserve the last surviving cow of the Enderby Island cattle breed. Reproduction, Fertility and Development 10: 369-378.

Wells, D.N.; Misica, P.M.; Day, A.M.; Peterson, A.J.; Tervit, H.R. 1999. Cloning sheep from cultured embryonic cells. Reproduction, Fertility and Development 10: 615-626.

Wells, D.N. 1999. Cloning livestock - heralding the new era of animal production. Livestock Improving Association of Japan, Embryo Transfer 21: 9-14.

McMillan, W.H.; Wells, D.N. 1999. Embryo and recipient contributions to the pregnancy loss following the transfer of cloned embryos derived from foetal and adult somatic cells. Proceedings of the New Zealand Society of Animal Production 59: 211-215.

L'Huillier, P.; Laible, G.; Wells, D. 1999. Genetic modification of livestock for the production of therapeutics and designer foods. Proceedings of the New Zealand Institute of Agricultural Science and the New Zealand Society for Horticultural Science Annual Convention: Food for Thought, June 30 - July 2 1999, pages 21-22.

Wells, D. 1999. Animal cloning provides many new opportunities for livestock production and biomedicine in the future. Agricultural Science 12 (3): 22-27.

Wells, D. 1999. The art of cloning. Dairy Exporter. October 1999, pages 90-92.

Wells, D.N. 1999. Animal cloning: current progress, challenges and future prospects. Proceedings of the XIII Brazilian Congress of Animal Reproduction, Revista Brasileria de Reproducao Animal 23: 86-97.

McMillan, W.H.; Wells, D.N.; Peterson, A.J.; Donnison, M.L. 1999. Early embryo development in nuclear transfer clones derived from somatic cells of a high genetic merit dairy cow. Association Europeene de Transfert Embryonnaire 15th Collogue Scientifique, Lyon 10-11 September, 1999, page 208.

Wells, D.N. 1999. Cloning for animal conservation and resurrection: facts and fantasies. Conference on Cloning the Huia: practical, ethical, and cultural issues. Hastings Boys' High School, July 9-10 1999.

Wells, D. 1999. Future opportunities in livestock production and biomedicine from advances in animal cloning technology. Proceedings of the New Zealand Association of Science Educators, BioLive Conference - Celebrating the Living World, Auckland, September 26-29. Abstract. Pages 13-14 and full paper.

Wells, D. 1999. Cloning and transgenesis in animals. Joint conference of the Animal Welfare Advisory Committee and the Australian and New Zealand Council for the Care of Animals in Research and Teaching, Wellington, November 18-19, Abstract, pp 16-17.

Wells, D. 1999. Nuclear transfer from established cell lines. Encylopaedia of Life Sciences www.els.net paper 5650.

Asher, G.W.; Berg, D.K.; Wells, D.N.; Scott, I.C. 2000. Genetic improvement and reproductive control of farmed red deer and wapiti (Cervus elaphus) - development and application of assisted reproductive technologies. Proceedings of the AAAP. In press.

L'Huillier, P.; Wells, D.; Laible, G. 2000. Development of genetically modified dairy cattle with enhanced milk casein levels. New Zealand BioScience. May 2000: 16-18.

Wells, D. 2000. Cloning and transgenesis in animals. Innovation, ethics, and animal welfare: public confidence in science and agriculture. Proceedings of the Joint conference of the Animal Welfare Advisory Committee and the Australian and New Zealand Council for the Care of Animals in Research and Teaching, Wellington, New Zealand, November 18-19, pages 88-96.

Wells, D. 2000. Somatic and embryonic cell cloning: what are the processes involved? In: Mid Northern Beef Council, Seminar for Beef Producers, Ruakura Research Centre, Hamilton, April 17 2000. pages 10-13.

Wells, D.N. 2000. Cloning and transgenesis in animals. New Zealand Pork Expo Conference, May 1 2000, Ellerslie, Auckland.

Wells, D.N. 2000. Mammalian cloning: techniques, applications and implications. In: Gene technology in New Zealand: scientific issues and implications, ERMA New Zealand Seminar, Te Papa, Wellington, May 6-7 2000. pages 13-15.

Wells, D.N. 2000. Livestock cloning and transgenesis (animal health and welfare aspects). New Zealand Veterinary Association Conference, Proceedings of the Industry Branch and the Food Safety & Biosecurity Branch, NZVA Auckland 8-11th June 2000, Publication no. 201: 19-27.

Wells, D.N. 2000. Cloning for animal conservation. Genetically Engineering and Cloning Animals: Science, Society and Industry, Park City, Utah, June 28-20 2000. Conference paper #3.

Piedrahita, J.A.; Miller, A.L.; Oliver, J.E.; Berg, M.C.; Forsyth, J.T.; Peterson, A.J.; Wells, D.N. 2001. Successful cloning in cattle with cytoplasts obtained from follicles of 1-3 mm in diameter. Theriogenology 55: 286.

Wrenzycki, C.; Wells, D.; Hermann, D.; Korsawe, K.; Hadeler, K.-G.; Miller, A.; Oliver, J.; Tervit, R.; Niemann, H. 2001. Effects of different activation protocols, cell cycle stages as well as passage numbers of donor cells on mRNA expression patterns in cloned bovine blastocysts. Theriogenology 55: 418.

Wrenzycki, C.; Wells, D.; Herrmann, D.; Miller, A.; Oliver, J.; Tervit, R.; Niemann, H. 2001. The nuclear transfer protocol affects mRNA expression patterns in cloned bovine blastocysts. Biology of Reproduction 65: 309-317.

McMillan, W.H.; Wells, D.N.; Peterson, A.J.; Donnison, M.L. 2001. Early embryo development in nuclear transfer clones derived from somatic cells of a high genetic merit dairy cow. British Society of Animal Science Occasional Publication 26 (part 1): 437-440.

Wells, D.; Miller, A.; Oliver, J.; Tucker, F.; Forsyth, J.; Berg, M.; Cockrem, K.; Oback, B.; Tervit, R. 2001. Greater post-natal viability with G0 compared to G1 donor cells following somatic cell nuclear transfer in cattle. Proceedings of the current status and perspectives in cloning and related studies. October 15-18 2001, Tsukuba, Japan, pp 73.

Wells, D.; Oliver, J.; Miller, A.; Forsyth, J.; Berg, M.; Cockrem, K.; Lange, J.; Tervit, R. 2001. Bovine cells selected in G1 of the cell cycle are totipotent following somatic cell nuclear transfer. Australian Society of Reproductive Biology. In press.

Laible, G.; Brophy, B.; L'Huillier, P.; Wells, D.N. 2001. Development of transgenic cattle with enhanced milk casein levels by nuclear transfer. Proceedings of Transgenic Animals in Agriculture Conference, Lake Tahoe USA. In press.

Laible, G.; L'Huillier, P.; Piedrahita, J.; Wells, D. 2001. Development of transgenic cattle for the production of human Myelin Basic Protein. BioScience. In press.

Wells, D.N.; Tucker, F.C.; Miller, A.L.; Oliver, J.E.; Xiang, T.; Forsyth, J.T.; Berg, M.C.; Cockrem, K.; Tervit, H.R. 2001. Effect of early or late G1 cell cycle phase on embryo development following somatic cell nuclear transfer in cattle. Theriogenology. In press.

Wells, D.N. 2001. The in vitro isolation of murine embryonic stem cells. In: Methods in Molecular Biology; Transgenesis Techniques: Principles and Protocols, Clarke A. (ed), Humana Press, Totawa, NJ. In press.

Wells, D.N. 2001. Production of chimaeras derived from murine embryonic stem cells. In: Methods in Molecular Biology; Transgenesis Techniques: Principles and Protocols, Clarke, A. (ed), Humana Press, Totawa, NJ. In press.

Steinborn, R.; Schinogl, P.; Wells, D.N.; Bergthaler, A.; Müller, M.; Brem, G. 2001. Interspecific heteroplasmy in somatic cattle clones. Proceedings of the National Academy of Sciences, USA. Submitted.

Oback, B.; Wells, D. 2001. Donor cells for cloning - many are called but few are chosen. Cloning and Stem Cells. Submitted.