2PN cell donation in Germany. Or: How the German Embryo Protection (Act) undermines itself

In contrast to embryo donation, the permissibility of 2PN cell donation is highly controversial in Germany. This article is based on there being a legal loophole with respect to 2PN cell donation, which results from an inconsistency within the Embryo Protection Act on the normative status of 2PN cells. Following that thesis, the article argues that, on the basis of the normative criterion totipotency (i.e. the capacity to develop into a born human being), 2PN cells should also be considered human embryos within the meaning of the Act and thereby be protected by that Act in the same way as embryos. However, the normative assumption that 2PN cells should already be endowed with human dignity and the right to life has absurd consequences. Moreover, the consistent continuation of the Embryo Protection Act, as well as of the underlying ethical position or argumentation (i.e. the potentiality argument), leads to the even more absurd consequence of having to place every human somatic cell under the protection of human dignity and the right to life. As totipotency or the developmental potential therefore cannot delimit entities considered worthy of protection (i.e. human embryos) from entities considered not worthy of protection (i.e. 2PN cells, gametes, hESC, hiPSC and human somatic cells), it is not a suitable normative criterion. As a paradigmatic case, 2PN cell donation demonstrates that by retaining this normative criterion the now obsolete German Embryo Protection (Act) ultimately undermines itself.     

鸡胚胎在生命科学研究中的应用

鸡作为优秀的模式动物在生命科学研究中的作用日益受到人们重视,其中以鸡胚胎的贡献尤为突出。本文简要综述了鸡胚胎在发育生物学、肿瘤、再生、神经生物学和胚胎干细胞等研究领域中的应用,同时对鸡基因组和鸡胚胎的研究方法也进行了简单探讨。     

Embryo Donation in Iran: An Ethical Review

Iran is the only Muslim country that has legislation on embryo donation, adopted in 2003. With an estimated 10–15% of couples in the country that are infertile, there are not any legal or religious barriers that prohibit an infertile couple from taking advantage of Assisted Reproductive Technologies (ARTs). Although all forms of ARTs available in Iran have been legitimized by religious authorities, there is a lack of legislation in all ARTs except embryo donation. By highlighting ethical issues in embryo donation, the paper presents a critical review of the Act of Embryo Donation in Iran. The paper argues that the Act does not provide enough safeguards for the future child and assurance for the safety of the donated embryos. It also does not restrict embryo donation to surplus embryos from infertile couples and is silent about the number of embryos that could be donated by each couple as well as the number of recipients for donated embryos by a couple. The Act is also silent about the issues of genetic linkage (nasab) and heritage which are challenging issues, especially in a conservative Islamic society. As a result, the future child may not inherit from their birth parents, as it is not required by the Act, or from the genetically related parents under the anonymity policy. Finally there is no standard national protocol or guidelines to evaluate the safety of the donated embryos. The paper concludes that despite its benefits, the Act lacks clarity, and it is subject to misunderstanding and confusion.     

NO ETHICAL BYPASS OF MORAL STATUS IN STEM CELL RESEARCH

Recent advances in reprogramming technology do not bypass the ethical challenge of embryo sacrifice. Induced pluripotent stem cell (iPS) research has been and almost certainly will continue to be conducted within the context of embryo sacrifice. If human embryos have moral status as human beings, then participation in iPS research renders one morally complicit in their destruction; if human embryos have moral status as mere precursors of human beings, then advocacy of iPS research policy that is inhibited by embryo sacrifice concerns renders one morally complicit in avoidable harms to persons. Steps may be taken to address these complicity concerns, but in the final analysis there is no alternative to achieving clarity with respect to the moral status of the human embryo.     

Regulating Consent to Organ and Embryo Donation The Legal Dimensions of the Problem

As rational adults, we are free to elect what is (or is not) done to our bodies. However, this strong freedom does not extend to the borders of life. Control over the uses of our biological material is constrained and uncertain at law. Our article examines the legal condition of embryos and organs: how law conceptualises them and regulates their uses.     

Prenatally diagnosed foetal malformations and termination of pregnancy: the case of Lebanon

Termination of pregnancy (TOP) is offered in many countries, for foetuses prenatally diagnosed with congenital malformations that are deemed incompatible with life or that are associated with a high morbidity. In Lebanon, a middle income country where religion plays a focal role, the law prohibits any form of TOP unless it is the only means to save the mother's life. It is the contention of the authors of this article that even if the foetus is a person, if it were medically revealed that there is a substantial risk that the newborn will suffer severe physical abnormalities that will cause it to be seriously handicapped; it is morally acceptable to terminate the pregnancy. Hence, TOP carried out for these indications is justified in the interest of the foetus and the child. Whatever the status of the foetus is, once born, it will become a full-fledged sentient being with all that this entails. When given the option of starting an existence, this person-to-be has the right to a minimum that allows him/her to enjoy a relatively good quality of life. Today, Lebanese obstetricians are confronted with the burden placed on them under the law to refuse TOP, or, when performing them, to forge records or deny having done them. This is why we strongly believe that the Lebanese policy on abortion should be amended.     

Framing the ethical and legal issues of human artificial gametes in research,therapy, and assisted reproduction: A German perspective

Recent results from studies on animals suggest that functional germ cells may be generated from human pluripotent stem cells, giving rise to three possibilities: research with these so‐called artificial gametes, including fertilization experiments in vitro; their use in vivo for therapy for the treatment of human infertility; and their use in assisted reproductive technologies in vitro. While the legal, philosophical, and ethical questions associated with these possibilities have been already discussed intensively in other countries, the debate in Germany is still at its beginning. A systematic and detailed analysis of the legal framework in Germany is provided with regard to the three possibilities, including the applicable statutory laws as well as the constitutional law. The question emerges as to whether the statutory laws as well as the constitution justify a distinction to be made between embryos of artificial and natural origin. This question is subject to philosophical analysis, discussing the distinction between person and thing, dignity and price, personality and property, and nature and technique. As a result, the criterion of naturalness alone may not be sufficient to differentiate between embryos of natural and artificial origin, and other criteria need to be identified.     

Chronology and dynamics of the development of female genital organs of cattle fetuses

The chronology and dynamics of the female germ cell development, of the mitotic activity of oogonia, and of the chromosome rearrangements at prophase I of meiosis have been quantitatively estimated in 30 cow embryos and foetuses at the age of 1.5 to 9 months. The sexual differentiation of the gonads was shown in a 1.5 month old embryo. The oocytes at the stages of preleptotene chromosome condensation and decondensation occurred in the 1.5 month old embryos and their maximum number was observed in the 2-5 month old foetuses. The leptotene oocytes were found in the 2-2.5 month old foetuses. The transition to zygotene and pachytene was also recorded in the 2-2.5 month old foetuses but their maximum number was observed in the 4-6 month old foetuses; their number was reduced to single oocytes thereafter. The first diplotene oocytes appeared in the 3 month old foetuses but the active transition of the oocytes to diplotene was observed after four months of development. The formation of a layer of follicle cells takes place around the diplotene oocytes. The vast majority of degenerating germ cells are the oocytes in zygotene-pachytene and in diplotene. The population of germ cells is formed by the mitotic division of oogonia in the cow foetuses, mainly at the age of 1.5 to 4 months of development.     

Efficient harvesting methods for early‐stage snake and turtle embryos

Reptile development is an intriguing research target for understating the unique morphogenesis of reptiles as well as the evolution of vertebrates. However, there are numerous difficulties associated with studying development in reptiles. The number of available reptile eggs is usually quite limited. In addition, the reptile embryo is tightly adhered to the eggshell, making it a challenge to isolate reptile embryos intact. Furthermore, there have been few reports describing efficient procedures for isolating intact embryos especially prior to pharyngula stage. Thus, the aim of this review is to present efficient procedures for obtaining early‐stage reptilian embryos intact. We first describe the method for isolating early‐stage embryos of the Japanese striped snake. This is the first detailed method for obtaining embryos prior to oviposition in oviparous snake species. Second, we describe an efficient strategy for isolating early‐stage embryos of the soft‐shelled turtle.     

鸡胚内抗氧化物质的分布与变化

生物体内存在多种内源性抗氧化物质,在生命过程中发挥着基本的防御功能,是人们十分关注的研究领域。本文综述了近年来鸡胚内抗氧化物质的形成与来源等研究成果,分析了鸡胚孵育过程中维生素(A、C、E)、类胡萝卜素、硒、过氧化氢酶和超氧化物歧化酶等主要抗氧化物质的分布与变化,及内源性抗氧化系统的形成,旨在为今后的研究提供有益的科学依据。     

Embryonic and foetal development of bovine oocytes treated with a combination of butyrolactone I and roscovitine in an enriched medium prior to IVM and IVF

Cattle oocytes were maintained at germinal vesicles (GV) stage for 24 hr using a combination of two specific and potent inhibitors of M-phase promoting factor (MPF) kinase activity, butyrolactone I (BL-I) and roscovitine (ROS). The media used for inhibition were (a) TCM-199 only and (b) TCM-199 supplemented with serum, hormones and growth factors. The effective doses of inhibitors were 6.25 microM BL-I and 12.5 microM ROS in medium (a) and 50 microM BL-I and 12.5 microM ROS in medium (b). After inhibition, about 90% of the oocytes resumed meiosis and reached the metaphase II (MII) stage during 24 hr of maturation. Following fertilisation the percentage of cleavage (D +2), compacted morula (D +6), blastocysts on D +7 and D +8 and the survival to freezing and thawing of grade 1 embryos frozen on D +7 were not different between the experimental treated groups and the control. In order to evaluate early foetal development, two groups of five grade 1 D +7 blastocysts derived from treated oocytes and two groups of five control embryos were transferred nonsurgically in four synchronised recipient heifers. On D +27, the recipients were slaughtered and the foetuses were recovered. In both groups, six foetuses developed out of the 10 embryos transferred. In conclusion, several supplements can be added to the prematuration medium of bovine oocytes without reducing the quality of inhibition but also without improving their subsequent developmental competence versus treated oocytes in TCM-199 only and versus untreated control. Furthermore, the prematuration step used in this study does not interfere with normal foetal development during the first stages of organogenesis.     

Experimentally produced diploid-triploid mouse chimaeras develop up to adulthood

Spontaneous diploid-triploid chimaeras occur sporadically in various mammalian species including man, but so far have never been produced experimentally. In order to get a deeper insight into the developmental consequences of this anomaly, we have developed two procedures that enabled for the first time to produce routinely diploid-triploid embryos, foetuses, and animals in the mouse. These procedures are: (1) aggregation of cleaving diploid embryos with triploid embryos produced by suppression of the second polar body in zygotes, and (2) fusion of a haploid karyoplast with one blastomere of the two-cell diploid embryos. The first procedure yielded 23 living and 6 dead postimplantation embryos and foetuses (age: 8th-19th day) out of which 22 were chimaeric. In addition, three chimaeric neonates reached adulthood. Two animals were fertile, and one--an overt chimaera--was an infertile male. The rate of postimplantation development of aggregation chimaeras was normal or only slightly retarded, and with one exception the foetuses were morphologically normal. Generally, the highest contribution of the 3n component in extra-embryonic structures was noted in the yolk sac, and usually it was higher than its contribution to the organs of the body. Chimaerism was most often noted in the liver, the heart, the intestine, and the lungs. Participation of triploid cells to all tissues studied, both in the body and in extra-embryonic structures, appeared to decrease slightly as development progressed. The second procedure yielded 10 foetuses and 6 adults. Three foetuses were chimaeric. Six fertile adults were probably non-chimaeras: the triploid component was absent in the coat and in the blood.     

Effect of sperm motility on human embryo quality in in vitro fertilization

Sperm motility is important for penetration of the zona pellucida, and this parameter has been reported to be the single most important factor determining fertilization rates. As there was no report on the relationship between sperm motility and embryo quality, we investigated the influence of sperm motility on embryo quality in 41 patients with tubal disease and/or obstruction. The patients were either unstimulated or stimulated with clomiphene or clomiphene and human menopausal gonadotrophin. Of 116 oocytes collected, 86 (74.1%) fertilised and cleaved; of these only 44 embryos had clear equal blastomeres without fragmentation (grade 3). Grade 3 embryos were equally distributed through all initial sperm motility categories, and through all categories of sperm concentration after swim-up. The ratio of motile sperm concentration in the initial semen sample to the final sperm concentration after swim-up varied from 0.5 to 67, and grade 3 embryos were distributed randomly from low to high ratios. The pregnancy rate in this series was only 14.6% per replacement. The rate of gestational sacs per embryo replaced was 7.0% (6/86); if “poor” embryos were excluded, the rate was 9.1% (6/66). The absence of correlation between sperm motility and embryo quality is discussed on morphological grounds.     

Active demethylation of individual genes in intracytoplasmic sperm injection rabbit embryos

Intracytoplasmic sperm injection (ICSI), as an assisted reproduction technique, has been widely used in animal and human. However, its possible effect on epigenetic changes has not been well studied. To investigate whether ICSI can induce aberrant DNA methylation changes in rabbit preimplantation embryos, we examined the methylation status of the SP-A promoter region and the satellite sequence Rsat IIE by bisulfite-sequencing technology. The SP-A promoter region was extensively demethylated before the first round of DNA replication commences, and the unmethylated status was maintained until morula when dynamic remethylation occurred. A similar but more moderate demethylation process was observed in satellite sequence Rsat IIE. These results are in contrast with the previous reports of no active demethylation in normal rabbit embryos, suggesting that the active demethylation we observed may be induced by ICSI.     

Protecting Future Children from In‐Utero Harm

The actions of pregnant women can cause harm to their future children. However, even if the possible harm is serious and likely to occur, the law will generally not intervene. A pregnant woman is an autonomous person who is entitled to make her own decisions. A fetus in‐utero has no legal right to protection. In striking contrast, the child, if born alive, may sue for injury in‐utero; and the child is entitled to be protected by being removed from her parents if necessary for her protection. Indeed, there is a legal obligation for health professionals to report suspected harm, and for authorities to protect the child's wellbeing. We ask whether such contradictory responses are justified. Should the law intervene where a pregnant woman's actions risk serious and preventable fetal injury? The argument for legal intervention to protect a fetus is sometimes linked to the concept of ‘fetal personhood’ and the moral status of the fetus. In this article we will suggest that even if the fetus is not regarded as a separate person, and does not have the legal or moral status of a child, indeed, even if the fetus is regarded as having no legal or moral status, there is an ethical and legal case for intervening to prevent serious harm to a future child. We examine the arguments for and against intervention on behalf of the future child, drawing on the example of excessive maternal alcohol intake.     

Birth of normal infants after transfer of embryos that were twice vitrified/warmed at cleavage stages: Report of two cases

The role of cryopreservation in assisted reproductive technology programs has increased within the last years allowing the transfer of a limited number of embryos and the storage of the remaining for future use. The reduction in the number of transferred embryos decreases the frequency of multiple pregnancy rates and of ovarian hyperstimulation syndrome while the cumulative pregnancy rate can be maximized. Moreover, as not all embryos will survive the warming process more cleavage stage embryos are warmed to improve selection for transfer. Therefore, surplus good quality cleavage stage embryos and/or blastocysts must be re-vitrified for further transfer to achieve pregnancy. To our knowledge, there have been no reports demonstrating that human embryos can be successfully vitrified/warmed twice at the cleavage stage. Thus we report two successful pregnancies and deliveries of healthy babies after transfer of embryos that were twice vitrified/warmed at 2–4 cells stage.     

Frozen embryos and the obligation to adopt

Rob Lovering has developed an interesting new critique of views that regard embryos as equally valuable as other human beings: the moral argument for frozen human embryo adoption. The argument is aimed at those who believe that the death of a frozen embryo is a very bad thing, and Lovering concludes that some who hold this view ought to prevent one of these deaths by adopting and gestating a frozen embryo. Contra Lovering, we show that there are far more effective strategies for preserving the lives of frozen embryos than adoption. Moreover, we point out that those who regard the deaths of frozen embryos as a very bad thing will generally regard the deaths of all embryos as a very bad thing, whether they are discarded embryos, aborted embryos or embryos that spontaneously abort. This entails that these other embryos must be taken into account when considering moral obligations, as well as other human lives at risk from preventable causes.     

Histological study of timing and embryology of notochordal abnormalities in rat exposed in utero to Doxorubicin

Experimental Doxorubicin-exposure in utero is correlated with foetal oesophageal atresia, tracheo-oesophageal fistula, axial alterations. While gastrointestinal and respiratory defects have been largely investigated, only sporadic data have been published to date on notochordal and vertebral defects. The aim of this work was the study of the genesis of chordal and vertebral abnormalities in rat embryos and foetuses exposed to Doxorubicin and the study of their correlation with oesophageal and tracheal defects. For this purpose, pregnant rats were i.p. injected with saline (control) or with 4 mg/Kg b.w. Doxorubicin on days 9.5 and 10.5 of gestation. Embryos and foetuses were morphologically analysed on days 10.5-15 and 16, 18, 20 of gestation respectively, fixed in formaldehyde and histologically processed. Slides were routinely stained with haematoxylin-eosin (11-15 days post coitum embryos and all foetuses) or specifically stained with aniline blue for the staining of basal laminae (10.5 days post coitum embryos). Moreover, some foetuses at term (20 days post coitum) were processed for bone and cartilage staining. The data obtained in the present work confirm the specificity of Doxorubicin in inducing gastro-intestinal and tracheal defects, describe the genesis of these defects step by step, describe the type and the genesis of notochordal abnormalities and their fate and exclude the role of Doxorubicin in inducing axial skeletal malformations.     

In vitro formation of tetraploid rat blastocysts after fusion of two-cell embryos

Gene targeting technology is not available in the rat which is an animal model of major importance, e.g., in cardiovascular research. This is due to the fact that the rat embryonic stem cell (ESC)-like cells established by several groups do not form germ-line chimeras when injected into blastocysts. In the mouse, the aggregation of ESC with tetraploid embryos has allowed the generation of animals completely derived from these cells. However, aggregation of rat ESC-like cells with tetraploid rat embryos has not yet been attempted to evaluate their developmental capacity. Therefore, we established a method to produce tetraploid rat embryos by fusion at the two-cell stage. Chemical fusion by polyethylene glycol (PEG) was shown to be less efficient (56.3% fused embryos) than electrofusion (96.1% fused embryos). The rate of development of fused embryos to blastocysts was independent of the fusion method and similar to the rate of control embryos. However, this rate was lower when the embryos had been cultured from the zygote state before fusion (14-20%) compared to freshly isolated two-cell embryos (41-63%). Alike for the mouse, blastocysts derived from fused two-cell rat embryos contained about half the number of cells as control blastocysts and were homogeneously tetraploid with no evidence of mosaicism. This method may be useful for the establishment of gene-targeting technology in the rat.     

Why Eberl is wrong. Reflections on the beginning of personhood

In a paper published in Bioethics, Jason Eberl has argued that early embryos are not persons and should not be granted the status possessed by them. Eberl bases this position upon the following claims: (1) The early embryo has a passive potentiality for development into a person. (2) The early embryo has not established both 'unique genetic identity' and 'ongoing ontological identity', which are necessary conditions for ensoulment. (3) The early embryo has a low probability of developing into a more developed embryo. This paper examines these claims. I argue against (1) that a plausible view is that the early embryo has an active potentiality to grow into a more developed embryo. Against (2), I argue that neither 'unique genetic identity' nor 'ongoing ontological identity' are necessary conditions for ensoulment, and that 'ongoing ontological identity' is established between early embryos and more developed embryos. Against (3), I argue that the fact that the early embryo has a low probability of developing into a more developed embryo, if true, does not warrant the conclusion that the early embryo is not a person. If Eberl is right that the human soul is that which organises the activities of a human being and that ensouled humans are persons, embryos are persons from conception.