Valoración de la adopción de embriones humanos congelados desde el punto de vista de la filosofía moral, la ética laica y dos religiones monoteístas / Valuating frozen human embryo adoption from a moral philosophical point of view, lay ethics and two monotheist religions / Valoração da adoção de embriões humanos congelados do ponto de vista da filosofia moral, a ética laica e duas religiões monoteístas

La búsqueda de la eficacia en la fecundación in vitro hace que se produzcan más embriones que los que se implantarán, lo que produce un excedente de embriones, que es congelado. Esto hace que ineludiblemente el número de embriones humanos congelados aumente. Entre las soluciones para dichos embriones humanos congelados está la donación/adopción de los mismos. Ineludiblemente esta práctica conlleva objetivos problemas éticos. En este trabajo se evalúa la eticidad de la donación/adopción de embriones humanos congelados desde la perspectiva de la filosofía moral, lo que podríamos llamar una "ética laica" y dos de las religiones monoteístas: la musulmana y la judía.

The search for IVF efficacy leads to a higher embryo production than it is necessary for implantation; this results in an excess of embryos which are kept frozen. This amount of frozen embryos inevitably increases. The donation/adoption are among the possible solutions for these frozen embryos. However, this practice has objective ethical problems. This article considers the ethical aspects of the donation / adoption of frozen human embryos from the point of view of moral philosophy, from what we could call "secular ethics" and from two monotheistic religions: Muslim and Jewish.

A busca da eficácia na fecundação in vitro faz com que se produzam mais embriões dos que se implantarão, o que produz um excedente de embriões, que é congelado. Isto faz com que inquestionavelmente o número de embriões humanos congelados aumente. Entre as soluções para os ditos embriões humanos congelados está na doação/adoção dos mesmos. Ineludivelmente esta prática implica objetivos problemas éticos. Neste trabalho se avalia a eticidade da doação/adoção de embriões humanos congelados a partir da perspectiva da filosofia moral, o que poderíamos chamar uma "ética laica" e duas religiões monoteistas: a mulçumana e a judia.

In vitro fertilization (IVF) in mammals: epigenetic and developmental alterations: scientific and bioethical implications for IVF in humans

The advent of in vitro fertilization (IVF) in animals and humans implies an extraordinary change in the environment where the beginning of a new organism takes place. In mammals fertilization occurs in the maternal oviduct, where there are unique conditions for guaranteeing the encounter of the gametes and the first stages of development of the embryo and thus its future. During this period a major epigenetic reprogramming takes place that is crucial for the normal fate of the embryo. This epigenetic reprogramming is very vulnerable to changes in environmental conditions such as the ones implied in IVF, including in vitro culture, nutrition, light, temperature, oxygen tension, embryo-maternal signaling, and the general absence of protection against foreign elements that could affect the stability of this process. The objective of this review is to update the impact of the various conditions inherent in the use of IVF on the epigenetic profile and outcomes of mammalian embryos, including superovulation, IVF technique, embryo culture and manipulation and absence of embryo-maternal signaling. It also covers the possible transgenerational inheritance of the epigenetic alterations associated with assisted reproductive technologies (ART), including its phenotypic consequences as is in the case of the large offspring syndrome (LOS). Finally, the important scientific and bioethical implications of the results found in animals are discussed in terms of the ART in humans.

Culture medium composition affects the gene expression pattern and in vitro development potential of bovine somatic cell nuclear transfer (SCNT) embryos

Different culture systems have been studied that support development of somatic cell nuclear transfer (SCNT) embryos up to the blastocyst stage. However, the use of sequential and two-step culture systems has been less studied. The objective of the present study was to examine the developmental potential and quality of bovine SCNT embryos cultured in different two-step culture media based on KSOM, SOF and the macromolecules FBS and BSA (K-K/FBS, K-S/BSA and K-K/BSA, respectively). No differences were observed in the cleavage rate for any of the culture systems. However, there was a significant difference (P<0.01) in the rate of blastocyst development, with the K-K/ FBS culture system yielding a higher rate of blastocysts (28%) compared to other treatments (18 and 15%, for K-S/BSA and K-K/BSA, respectively). Although quality of embryos, as assessed by the total number of cells, was not different, the apoptosis index was significantly affected in the sequential culture system (K-S/BSA). Gene expression analysis showed alterations of DNMT1, IGF2, LIF, and PRDX6 genes in embryos cultured in K-S/FBS and of SOD2 in embryos cultured in K-K/BSA. In conclusion, we demonstrated that culture medium may affect not only the developmental potential of SCNT embryos but also, more importantly, the gene expression pattern and apoptotic index, presenting the possibility to manipulate the culture medium composition to modulate global gene expression and improve the overall efficiency of this technique.

Análisis de patrones morfológicos y anatómicos en la embriogénesis somática del banano Williams (AAA) / Analysis of the morphological and anatomical patterns in the somatic embryogenesis of Williams banana (AAA)

La embriogénesis somática representa una herramienta esencial en el mejoramiento genético y en la micropropagación clonal masiva de bananos mejorados. En el presente trabajo se analizaron los patrones morfológicos y anatómicos que ocurren durante la embriogénesis somática del banano Williams, dirigidos a conocer y mejorar este proceso. En la investigación se establecieron suspensiones celulares embriogénicas (SCE) a partir de callo embriogénico obtenido de manos florales inmaduras masculinas, las cuales originaron abundantes embriones que regeneraron plantas. Hacia los tres meses de cultivo se detectaron embriones somáticos (ES) primarios color blanco-crema en las manos florales de los nudos nueve a doce, contados a partir del ápice floral. Al cuarto mes estos ES primarios dieron origen al callo embriogénico, de color blanco crema, estructura granular, con abundantes ES torpedo en su periferia y con una organización celular en tres diferentes zonas. De este callo se cultivaron porciones pequeñas con ES torpedo en medio de multiplicación durante dos meses, dando origen a la SCE I. La misma se tamizó (250 µm) para establecer la SCE II. El sedimento de células y los agregados celulares embriogénicos de ambas SCE se trasladó a medio de maduración. Transcurridos dos meses los embriones maduros se transfirieron a medio de conversión de embriones, lográndose regenerar plantas completas a partir de las dos semanas. Las SCE produjeron numerosos embriones somáticos maduros y mostraron una buena conversión de embriones a plantas y regeneración de plantas. Este sistema de embriogénesis somática permitió la obtención de plantas funcionales en nueve meses.

Somatic embryogenesis represents an essential tool for the genetic improvement and for the mass clonal micropropagation of the improved banana plant. In this present work morphological and anatomical patterns were analyzed in the somatic embryogenesis of Williams banana, to know and enhance this process. In the investigation embryogenic cell suspensions (ECS) were established from embryogenic callus obtained from floral immature male hands, which gave rise to many somatic embryos that regenerated plants. Towards the three months of culture white-cream primary somatic embryos (SE) were detected in the floral hands of the nodes nine to twelve, counted from the floral apex. At the fourth month this primary SE gave origin to a creamy-white embryogenic callus, with granular structure and abundant SE torpedo on its periphery. Cell organization with three different zones was observed in callus. Small portions of this callus were cultivated in the multiplication medium for two months, to originate ECS I. This ECS was filtered through a mesh (250 µm pore size) to establish the ECS II. The sediment of embryogenic cells and cell clusters of the ECS were moved to maturation media. After two months the mature embryos were transferred to conversion medium, and two weeks later, whole plants were developed. The ECS produced numerous mature SE, which showed good conversion of embryos into plants and plant regeneration. This system of somatic embryogenesis permitted the mass production of functional plants in nine months.

Assessment of in vitro maturation and fertilization of BDF1 mice germinal vesicle oocytes with and without cumulus cell after vitrification

Improving pregnancy rate associated with the use of cryopreserved oocytes would be an important advancement in human Assisted Reproductive Technology [ART]. Vitrification allows glasslike solidification of a solution, a physical process, without ice crystal formation in the living cells. The purpose of this study was to evaluate the viability of the oocytes, in vitro maturation and embryo development vitrified germinal vesicle oocytes after single and stepwise exposure to cryoprotectants. Germinal vesicle oocytes or without cumulus cells were transferred to a verification solution composed of 30% M sucrose [v/v] ethylene glycol, 18% [w/v] Ficoll-70, and 0.3 M sucrose either by single step or in a step-wise fashion. After verification and storage in liquid nitrogen, the oocytes were thawed and washed twice in the medium TCM 119 and then subjected to in vitro maturation, the capacity of fertilization and embryonic development to 2-cell were examined in vitro. The oocytes surviving, maturing to MII, fertilization developmental rate in the step-wise exposure were significantly higher [P<0/05], compared with the corresponding rate in single step procedure. The results of the present study indicated that oocytes vitrified with cumulus cells and stepwise procedure had a positive effect on the maturation and developmental rate than oocytes without cumulus cells and single step procedure

Modulation of murine blastocyst hatching in vitro by glutamine and tryptophan

Enrichment of culture media with amino acids improves embryo development. However, little is known about the specific action of each amino acid during embryogenesis. The present study was undertaken to examine the effect of L-glutamine (Gln) and tryptophan (Trp) on mouse embryo hatching, expansion and viability in vitro. Blastocysts were collected from 6- to 8-week-old female BALB/c mice (N = 30) and cultured in M2 medium containing either 0.125, 0.25 or 0.5 mM Trp, 1 mM Gln, or M2 alone. Gln significantly increased (100 percent; P < 0.05) blastocyst hatching at 24 h compared to M2 alone or Trp; moreover, Trp inhibited blastocyst hatching when compared to M2 alone (P < 0.05) at 72 h. In contrast, the percentage of embryos reaching the state of expanded blastocyst at 48 h was significantly higher in medium with 1 mM Gln (66.6 percent; P < 0.05) or with 0.125 mM Trp (61.1 percent; P < 0.05). Unexpectedly, Trp increased the percentage of degenerated blastocysts after 48 h (67.7 percent; P < 0.05), while Gln preserved blastocyst viability. These results suggest that Gln may enhance blastocyst hatching, expansion and viability in vitro.

Extended culture up to the blastocyst stage: a strategy to avoid multiple pregnancies in assisted reproductive technologies

The aim of this study was to review the experience and outcomes of assisted reproduction cycles with embryos grown up to day 5 of development, comparing different parameters according to the ages of the patients. We retrospectively studied 1,874 assisted reproduction cycles where embryo culture was extended up to the fifth or sixth day of development. All IVF and ICSI cycles were included, comparing, according to patient age, the following rates: blastocyst formation, pregnancy, implantation and abortion. As control, we analyzed cycles with donated oocytes from young donors (OD). The number of embryos reaching the blastocyst stage is similar in all groups of patients. Only the OD group was different in terms of blastocyst formation, pregnancy and implantation rates. Patients over 39 years of age had an abortion rate of 59.1 percent, which is significantly higher than the other groups. Extended embryo culture up to the blastocyst stage can be implemented in programs of assisted reproduction in order to increase the pregnancy rate. The potential of blastocyst implantation is high, allowing us to transfer fewer embryos and reduce the probability of multiple pregnancies.

Fertilização in vitro: mais de 4 milhões de crianças nascidas e um Prêmio Nobel / In vitro fertilization: more than 4 million children born and a Nobel Prize

Em 1978, nasceu Louise Brown, o primeiro bebê de proveta. Este nascimento histórico foi o resultado de pesquisas do biólogo Robert Edwards e do ginecologista Patrick Steptoe (1). Em 2010, Edwards recebeu o prêmio Nobel de Medicina/Fisiologia, pela importância desta técnica para a humanidade. Este artigo revisa as etapas envolvidas na fertilização in vitro (FIV), as dificuldades dos pioneiros, as soluções encontradas e os procedimentos clínicos utilizados atualmente.

Louise Brown, the first baby conceived by in vitro fertilization (IVF), was born in 1978. This landmark was a result of research conducted by biologist Robert Edwards and gynecologist Patrick Steptoe (1). In 2010, Edwards was awarded the Nobel Prize in Physiology or Medicine in recognition of the importance of IVF to humanity. The objective of the present article is to review the steps involved in IVF, the difficulties faced by pioneers, the solutions found, and the clinical procedures currently employed.

Falha na sexagem por inibição do desenvolvimento de embriões bovinos produzidos in vitro com anticorpos anti H-Y / Failure of sexing by developmental arrest of bovine embryos in vitro produced with H-Y antisera

Embriões bovinos produzidos in vitro, em estádio de mórula, foram cultivados em meio contendo anticorpos anti H-Y de alto título proveniente de ratos por 24h e, após este tempo, classificados em dois grupos: 1) embriões inibidos em estádio de mórula (classificados como machos) e 2) embriões que se desenvolveram e formaram a blastocele (classificados como fêmeas). O sexo de 311 embriões, distribuídos em três grupos de concentração dos anticorpos, 3 por cento, 5 por cento ou 7 por cento, foi identificado pela reação em cadeia da polimerase. Não houve desvio da proporção entre machos e fêmeas (P>0,05) nos grupos em que se utilizaram os anticorpos anti H-Y, quando comparadas ao grupo-controle, sem adição de anticorpos anti H-Y. Diferentemente dos resultados obtidos utilizando-se embriões bovinos produzidos in vivo, a sexagem com anticorpos anti H-Y de alto título em embriões produzidos in vitro não propiciou sucesso.

In vitro produced bovine embryos at morula stage were cultured in medium containing high titer of rat H-Y antisera for 24h. The embryos were classified in two groups: 1) embryos arrested at morula stage (classified as males); and 2) embryos that developed and formed a blastocoele (classified as female). The sex of 311 embryos, divided in three groups of concentration of H-Y antisera, 3 percent, 5 percent or 7 percent, was identified by polimerase chain reaction. The results showed no difference (P>0.05) on sexual deviation in groups in which the H-Y antisera was added, in relation to control group, in which no H-Y antisera was added. In contrast with results obtained with in vivo produced bovine embryos, the sexing of in vitro produced bovine embryos with high H-Y antisera titer did not succed.

Inter-specific hybridization between freshwater catfish Mystus cavasius (Ham & Buch) and M. seenghala (Sykes) by artificial fertilization.

By employing the technique of induced ovulation for artificial fertilization, inter-specific hybrids between the threatened catfish species (Mystus cavasius x M. seenghala) were produced. Fertilization, hatching and survival were significantly different between control and hybrids. The hatching time of the hybrid was significantly lower than that of the control. The average performance viz., hatching time and viability of larvae of the control fish, was significantly better than that of the hybrids. In the hybrid cross, hatchlings were mostly deformed and abnormal and after yolk absorption ultimately succumbed.