Opinion of the Group of Advisers on the Ethical Implications of Biotechnology of the European Commission

The following points aim to shed light on the cloning debate by giving information on the scientific aspects of cloning and the ethical problems relating to them.

1.1. Cloning is the process of producing "genetically identical" organisms. It may involve division of a single embryo, in which case both the nuclear genes and the small number of mitochondrial genes would be "identical," or it may involve nuclear transfer, in which case only the nuclear genes would be "identical." But genes may be mutated or lost during the development of the individual: the gene set may be identical, but it is unlikely that the genes themselves would ever be totally identical. In the present context, we use the term "ge netically identical" to mean "sharing the same nuclear gene set."

1.2. It is inherent in the process of sexual reproduction that the progeny differ genetically from one another. In contrast, asexual reproduction (cloning) produces genetically identical progeny. This is a common form of reproduction in plants, both in nature and in the hands of plant breeders and horticulturists. Once a desired combination of characteristics has been achieved, asexual reproduction is the best way of preserving it. Asexual reproduction is also common among some invertebrate animals (worms, insects). Asexual reproduction in plants and invertebrates usually takes place by budding or splitting.

1.3. The first successful cloning in vertebrate animals was reported in 1952, in frogs. Nuclei from early frog embryos were transferred to unfertilized frog eggs from which the original nuclei had been removed. The resulting clones were not reared beyond the tadpole stage. In the 1960s, clones of adult frogs were produced by transfer not only of nuclei from early embryos but also of nuclei from differentiated larval intestinal cells. Later, clones of feeding tadpoles were obtained by nuclear transfer from differentiated adult cells, establishing that differentiation of cells involving selective gene expression does not require the loss or irreversible inactivation of genes. Nuclear transfer in frogs has not yet generated an adult animal from cells of an adult animal.

1.4. Nuclear transfer can be used for different objectives. Nuclear transfer in mice has been used to show that both a female and a male set of genes are required for development to birth. If the two pronuclei, taken from fertilized eggs and transferred into an enucleated egg, are only maternal or only paternal, normal development does not occur. This is not cloning, since the single embryo formed is not identical to any other embryo and the objective is not to multiply individuals.

1.5. Nuclear transfer has also been used for cloning in various mammalian species (mice, rabbits, sheep, cattle), but until recently only nuclei taken from very early embryos were effective, and development was often abnormal, for reasons that are not fully understood.

1.6. In contrast, cloning by embryo splitting, from the 2-cell up to the blastocyst stage, has been extensively used in sheep and cattle to increase the yield of progeny from genetically high-grade parents. Because of the different pattern of early development, embryo splitting is much less successful in mice. From a scientific point of view, it would probably not be very effective in the human, although monozygotic (one-egg) twins and higher multiples occur naturally at a low incidence.

1.7. In 1996, a new method of cloning sheep embryos was reported, which involved first establishing cell cultures from single embryos. Nuclei from the cultured cells were transferred to enucleated unfertilized sheep eggs, particular attention being paid to the cell cycle stage of both donor and host cells, and the eggs were then artificially stimulated to develop. Genetically identical normal lambs were born.

1.8. Cell cultures were then established not only from embryonic and fetal stages, but also from mammary tissue taken from a 6-year-old sheep. Nuclear transfer was carried out as before, and in 1997 it was reported that several lambs had been born from the embryonic and fetal transfers and one lamb named Dolly (out of 277 attempts) from the adult nuclear transfer. It is not known whether the transferred nucleus was from a differentiated mammary gland cell or from a stem cell.

1.9. From the point of view of basic research, this result is important. If repeatable it may allow greater insight into the aging process, how much is due to cell aging, and whether or not it is reversible. Such work may also increase our understanding of cell commitment, the origin of the cancer process, and whether it can be reversed, but at the present time the research is at a very early stage. Dolly may have a shortened life span or a greater susceptibility to cancer: if she is fertile, her progeny may show an increased abnormality rate, owing to the accumulation of somatic mutations and chromosomal damage.

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