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PubMed1979

Naegleria fowleri in chick embryos. Effects of embryo age and incubation temperature, and the infectivity of embryo-derived amebae for mice.

Chick embryos were infected with Naegleria fowleri which was initially isolated from an ultimately fatal human case. Following inoculation of equivalent numbers of amebae on the chorioallantoic membrane, younger embryos died earlier than older embryos infected at the same time. Incubation of infected embryos at 32 degrees C prolonged survival only slightly in comparison with those at 37 degrees C. N. fowleri maintained for more than 25 serial passages in chick embryos retained infectivity for mice and the ability to convert to the biflagellate form in vitro.

Age Factors↗
PubMed1976

Wheat embryo ribonucleates. VI. Comparison of the 3'-hydroxyl termini in 'rapidly labelled' RNA from metabolizing wheat embryos with the corresponding termini in ribosomal RNA from differentiating embryos of wheat, barley, corn and pea.

The NaCl-insoluble (2.5 M, 0 degrees C) fraction of wheat embryo RNA (iRNA) can be labelled when wheat embryos are subjected to either short-term (0.5 h) or long-term (24 h) imbibition in a medium that contains tritium-labelled adenosine, guanosine, cytidine and uridine. Electrophoretic analyses reveal that, after short-term labelling, there is a broadly heterodisperse distribution of radioactivity in 'rapidly labelled' i[3H]RNA, but after long-term labelling, there is an essentially trimodal distribution of radioactivity in i[3H]RNA. End-group analyses reveal that, after short-term labelling, adenosine is the principal 3'-hydroxyl terminus in all centrifugal subfractions of 'rapidly labelled' i[3H]RNA, whereas cytidine (in 5.8S rRNA), guanosine (in 18S rRNA) and uridine (in 26S rRNA) are the principal 3'-hydroxyl termini in centrifugal subfractions of wheat embryo i[3H]RNA. Guanosine is also the principal 3'-hydroxyl terminus in the 18S rRNA of differentiating embryos excized from both monocotyledonous (wheat, barley, corn) and dicotyledonous (pea) seedlings. The implications that the end-group measurements may have for current views about the possible biochemical involvements of 3'-hydroxyl terminal sequences in both mRNA and 18SrRNA are subjects of discussion. Incidental to the principal investigation, an existing technique for analyzing the RNA contents of cellular materials has been appropriately modified to circumvent interference from uv-absorbing pigments, which, when present, prevent application of the method to plant materials.

Base Sequence↗
PubMed1994

The triumph of the pre-embryo: interpretations of the human embryo in Parliamentary debate over embryo research.

In 1984, both British Houses of Parliament overwhelmingly condemned the proposal in the Warnock Report that research be allowed on human embryos, under licence, during the first two weeks of their existence. In 1990, legislation permitting such research and based directly on the Warnock proposal was approved with substantial majorities in the two Houses. This study describes some of the cultural developments underlying the swing within Parliament from almost total rejection of embryo research to eventual acceptance. Material taken from the parliamentary record and from the Warnock Report is used to illustrate how two competing images of the human embryo provided a central focus for the adversarial process whereby legislation on embryo research came to be enacted.

Embryology↗
PubMed1999

A prospective randomized study comparing the outcome of in-vitro fertilization and embryo transfer following culture of human embryos individually or in groups before embryo transfer on day 2.

A prospective randomized trial of in-vitro fertilization and embryo transfer was undertaken to investigate the reported beneficial effects of culturing preimplantation human embryos in groups, rather than individually. A total of 159 treatment cycles, in which the women were matched for age, basal gonadotrophin concentrations and number of previous attempts, were included in the study. Of these, 78 cycles were randomized to the 'individual culture' group, and 81 cycles were randomized to the 'group culture' group. The groups did not differ in terms of the median number of oocytes or embryos obtained per cycle. There was no statistically significant difference between the two groups in terms of treatment outcome, as assessed by pregnancies or clinical pregnancies.

Adult↗
PubMed1996

Incidence of microbial growth from the tip of the embryo transfer catheter after embryo transfer in relation to clinical pregnancy rate following in-vitro fertilization and embryo transfer.

A total of 110 consecutive women was studied prospectively at the time of transcervical embryo transfer following conventional in-vitro fertilization and intracytoplasmic sperm injection procedures. Microbiological cultures were performed on endocervical swabs and embryo transfer catheter tips. Positive microbial growths were observed from endocervical swabs in 78 (70.9%) women and from catheter tips in 54 (49.1%) women. The clinical pregnancy rates were 57.1% in the group of patients without growth and 29.6% in the group with positive microbial growth from catheter tips. As microbial contamination at embryo transfer may influence implantation rates, prospective studies are justified to determine whether eradication of endocervical micro-organisms is possible and whether their eradication will improve implantation rates.

Adult↗
PubMed1999

Pattern of Brachyury gene expression in starfish embryos resembles that of hemichordate embryos but not of sea urchin embryos.

Echinoderms, hemichordates and chordates are deuterostomes and share a number of developmental features. The Brachyury gene is responsible for formation of the notochord, the most defining feature of chordates, and thus may be a key to understanding the origin and evolution of the chordates. Previous studies have shown that the ascidian Brachyury (As-T and Ci-Bra) is expressed in the notochord and that a sea urchin Brachyury (HpTa) is expressed in the secondary mesenchyme founder cells. A recent study by [Tagawa et al. (1998)], however, revealed that a hemichordate Brachyury (PfBra) is expressed in a novel pattern in an archenteron invagination region and a stomodaeum invagination region in the gastrula. The present study demonstrated that the expression pattern of Brachyury (ApBra) of starfish embryos resembles that of PfBra in hemichordate embryos but not of HpTa in sea urchin embryos. Namely, ApBra is expressed in an archenteron invagination region and a stomodaeum invagination region.

Amino Acid Sequence↗
PubMed1998

Statistical models predicting embryo survival to term in cattle after embryo transfer.

Embryo survival to term in recipient cattle is highly variable. We examined calving data in the published literature to determine whether a model of binomial independence or a model which includes an embryo (e) and recipient term (r), adequately explain observed embryo survival rates following attempts to induce twin calving using transfer of two embryos. To achieve this we examined 32 published papers which provided us with 47 sets of data concerning 4560 recipients with either 0, 1 or 2 calves born. In each set of data, the observed embryo survival rate to term (p) (number of calves born/number of embryos) was calculated and the expected number of recipients with either 0, 1 or 2 calves born was determined, assuming a binomial distribution. Parameters for the second model were estimated using maximum-likelihood procedures. The model of embryo independence was rejected in 85% of the sets of data, suggesting that factors other than the embryo are important sources of variation in embryo survival or loss. The proposed e and r model of embryo survival adequately describes the published data in recipients receiving either single or twin embryos. In general, only 50-70% of embryos and recipients are sufficiently competent to result in a calving. Variation among laboratories producing either in vitro or in vivo derived embryos was due to variation in recipient and not embryo competence. It is argued that e rather than observed embryo survival rate, and r rather than observed pregnancy rate, should be used to compare differences among embryo treatments and groups of recipients, respectively. Acceptance of this proposition should permit faster progress in identifying the biology of superior embryos and recipients, which is a prerequisite to improving embryo survival rate in cattle. Collectively, the published data are not consistent with a model of embryo independence, and that a model of embryo survival to term which recognises recipient as well as embryo contributions to embryo survival may be more appropriate in cattle.

Animals↗
PubMed1995

Embryo score to predict implantation after in-vitro fertilization: based on 957 single embryo transfers.

The purpose of this study was to devise an embryo score to predict the likelihood of successful implantation after in-vitro fertilization (IVF). Unlike most studies dealing with the influence of embryo stage and morphology on pregnancy, our study was based on single rather than multiple embryo transfers. A total of 957 single embryo transfers were carried out. No delivery was obtained after any of the 99 transfers using 1-cell embryos or embryos obtained after delayed fertilization. In the remaining 858 transfers, the embryos had cleaved. Higher pregnancy rates were obtained with embryos displaying no irregular cells (11.7 versus 6.9%; P < 0.01) and embryos displaying no fragmentation (11.5 versus 8.1%; P < 0.05). The 4-cell embryos implanted 2-fold more often than embryos with more or less cells (15.6 versus 7.4%; P < 0.01). Based on these observations, we devised a 4-point embryo score in which embryos are assigned 1 point each if they (i) are cleaved, (ii) present no fragmentation, (iii) display no irregularities, and (iv) have four cells. Both pregnancy rate and take home baby rate were significantly correlated with embryo score. Each point of this score corresponds to a 4% increase in pregnancy rate. Interestingly, pregnancy rate was significantly lower in women aged > 38 years (8.2 versus 11.4%; P < 0.05), even though embryo quality was similar regardless of age. Single embryo transfer allowed us to define a simple and useful embryo score to choose the best embryo for transfer to optimize IVF and embryo transfer outcome. The use of this embryo score could decrease multiple pregnancies after multiple embryo transfers.

Adult↗
PubMed2002

Developments in the storage of embryos in France and the limitations of the laws of bioethics. Analysis of procedures in 17 storage centres and the destiny of stored embryos.

BACKGROUND TO THE STUDY: 1985 witnessed the first transfers of frozen embryos resulting in live births in France. Since this time the number of embryos obtained by in vitro fertilisation (IVF) has increased each year. In 1999 each IVF attempt obtains, on average, 4.5 embryos that can be successfully implanted. In this paper we consider only those couples who have successfully obtained embryos (either by ICSI or traditional IVF techniques). The aims of the study are: To show how developments in embryo production and conservation have influenced the number of embryos stored. To address the socio-medical and ethical issues raised and to provide practitioners with some thoughts for reflection when consulting with couples based on the study findings To discuss the results of our findings in the light of those ethical questions raised by the imminent revision of the Laws of Bioethics. METHOD: In the first instance we did a retrospective analysis of quantitative data that 17 storage centres had collected over a period of 5 years. This period was marked by the implementation in 1994 of Laws described as Bioethics' Laws in France. During a second period we conducted a qualitative study regarding the fate of stored embryos. In order to do this, we began an analysis of the "status" of embryos and the decisions of those couples whose embryos were still in storage. For this a questionnaire was used. FINDINGS: The number of embryos that remain in storage in the 17 storage centres has increased reaching a total of 17,592 embryos involving 3,888 couples. The results show a consistent and persistent increase in the number of embryos stored before and after 1994. The qualitative study shows that: 51% of couples with embryos in storage can no longer be found, 23.6% request a continuance of storage, 12% would accept donating their embryos to medical research, 9.1% would wish for other couples to take eventual ownership of the embryo in 7.2% of cases the storage centre has can provide no information concerning the continuing of storage of such embryos. INTERPRETATION: The Bioethics Laws have therefore not succeeded in limiting the inflation in the number of embryos stored despite that the fact that this was one of the major concerns of those involved in formulating the laws and the medical professionals involved. Our study shows that the guidelines provided by these Laws remain ambiguous and that the objectives defined therein are thus difficult to achieve. Our study highlights the impact of such developments on the possible eventual "destiny" of such embryos, the behaviour of and the advice provided to couples, and the management practice of the storage centres involved. We present these results in the light of the laws established in 1994 in order to define not only the benefits but also the potential limitations of such legislation. No guidelines were laid down regarding the future of embryos belonging to a couple that no longer wishes to embark upon a "parental project". As a result it is not possible to terminate the conservation of such embryos which therefore remain in storage awaiting the revision of the current laws. The major objectives of our study assess the impact of the Laws and their impact on practice and to contribute to the debate which has yet to take place in the social and political arena.

Biomedical Research↗
PubMed2004

[Effect of early cleavage of embryos after intracytoplasmic sperm injection (ICSI) into oocytes on results of fertilization in vitro (IVF) and embryo transfer (ET)].

OBJECTIVE: To evaluate the influence of early cleavage in embryos obtained using ICSI on the success rate of IVF-ET. DESIGN: A prospective study. SETTING: Centre of Assisted Reproduction, Department of Obstetrics and Gynecology, Medical Faculty, Palacký University, Olomouc. METHODS: The rate of the first cleavage of embryos was checked between 23 to 26 hours after ICSI in 115 cycles IVF-ET. "Early Cleavage" (EC) embryos which were already cleaved were cultivated separately from the "slow" (No Early Cleavage, NEC) embryos. After 3 days cultivation, no more than three 6- to 8-cell embryos were transferred. Only EC embryos were transferred in 43 cycles and only NEC embryos were transferred also in 43 cycles. The remaining 29 cycles, where both EC and NEC embryos had to be transferred, were not evaluated. In both groups the following entry parameters were compared: age of the patient, number of fertilized oocytes, fertilization rate (FR), number of frozen and evaluated embryos, number of EC embryos recorded and number of transferred embryos. The outcome parameters compared were: pregnancy rate (PR), number of gestational sacs with heart beats per number of transferred embryos (implantation rate, IR), abortions (AB) and the frequency of multiple pregnancy. Statistical evaluation was performed by t-test, chi 2 test and by Sheffe analysis. RESULTS: EC embryos were found in 57% of cycles (n = 66) after ICSI. Early cleavage was observed in 198 of 563 embryos (35%). Among entry parameters the only statistically significant differences (P < 0.05) were found in the number of retrieved oocytes as well as in the ratio of frozen embryos which were higher in the cycles with the transfer of only EC embryos. FR was the same (74%) in both types of transfer. Outcome parameters in the transfer of only EC versus NEC embryos were higher in PR (40% vs. 33%), about equal in IR (27% vs. 25%) and lower in AB (2% vs. 5%). Moreover, multiple pregnancies were less frequent in the transfer of only EC embryos: 65% of singleton pregnancy, 35% of twins and no triplet versus 36%, 57% and 7%, resp., in the transfer of only NEC embryos. The differences in outcome parameters are not statistically significant (P > 0.05) because of small numbers. CONCLUSION: The speed of the first cell cleavage is a useful additional criterion for better evaluation of quality and viability of embryos.

Adult↗
PubMed2005

Embryo fragmentation as a determinant of blastocyst development in vitro and pregnancy outcomes following embryo transfer.

OBJECTIVE(S): To determine how the type of embryo fragmentation on day 3 affects progression of human embryos to blastocyst and pregnancy rates following embryo transfer. STUDY DESIGN: Retrospective analysis of all in vitro fertilization cycles in patients < or =40 years of age or younger from January 2002 through December 2003, during which time surplus day 3 embryos were transferred to blastocyst medium for extended culture. All embryos (4 cells or more) not suitable for transfer or freezing 72 hours following in vitro fertilization were placed into microdroplets (60 microL) of blastocyst medium and cultured for an additional 48 hours to assess blastocyst formation. Normal blastocyst development required blastulation, a visible inner-cell mass, trophectoderm cells covering 60% of the inner zona surface and thinning of the zona. The rate of blastocyst formation was then analyzed (chi 2 and analysis of variance) against the type of fragmentation 72 hours after insemination. Pregnancy outcomes were analyzed with respect to the pattern of fragmentation in cleaving embryos transferred after 3 days of culture. RESULTS: A total of 1566 embryos were cultured beyond day 3 of development of which 229 (14.6%) reached the blastocyst stage and were frozen. Embryos exhibiting no fragmentation or type I fragmentation had significantly higher blastocyst development rates (27.9% and 19.9%) than embryos with type 2 or 3 fragmentation (13.9 and 8.8, respectively; P < .001). No embryos with type 4 or 5 fragmentation progressed to blastocyst. The average type of fragmentation in transferred embryos correlated with pregnancy outcome and embryo age. CONCLUSION(S): More pervasive embryo fragmentation was associated with a decreasing rate of blastocyst development with day 3 embryos. To the extent that blastocyst development rates of day 3 embryos is an index of embryo viability, our findings establish that careful classification of the type of embryo fragmentation is important in selection of day 3 embryos for transfer. Recent reports of associations among embryo fragmentation, aneuploidy, apoptosis, and patient age support these conclusions.

Adult↗
PubMed1986

An approach to successful freezing of demi-embryos derived from day-7 bovine embryos.

The developmental capacity of frozen/thawed bisected embryos (n = 33) derived from day-7 bovine embryos was investigated and compared to ordinary embryos after freezing and thawing (n = 28) and to freshly bisected embryos (n = 19). The freezing and thawing protocol was identical for ordinary and demi-embryos. The percentage of intact embryos classified as excellent, good, or poor after thawing was 92.9 and 96.3% for ordinary and demi-embryos, respectively. Pregnancy rates of 53.8 (8 15 ), 46.2 (6 13 ), and 47.5% (9 19 ) were obtained when frozen/thawed ordinary embryos and frozen/thawed demi-embryos classified as excellent or good and sealed with an additional zona pellucida from hatched pig blastocysts or freshly bisected embryos were transferred. One pair of identical twins resulted from the transfer of frozen/thawed demi-embryos sealed with an additional zona pellucida. Transfer of four frozen/thawed demi-embryos without an additional zona pellucida led to one pregnancy. In contrast, demi-embryos derived from frozen/thawed ordinary embryos (n = 8) as well as frozen/thawed demi-embryos classified as poor (n = 6) did not result in any pregnancies although two halves were transferred per recipient. It is concluded that sealing the punctured zona pellucida improves the developmental capacity of frozen/thawed demi-embryos derived from day-7 bovine embryos, and freezing demi-embryos is more efficient compared to the splitting of frozen/thawed ordinary embryos.

Journal Article↗
PubMed2005

Relationship between pre-embryo pronuclear morphology (zygote score) and standard day 2 or 3 embryo morphology with regard to assisted reproductive technique outcomes.

OBJECTIVE: To test the hypothesis that pregnancy rates are low if grade Z1 pre-embryos are not available for transfer and to determine if pronuclear morphology is a better predictor of pregnancy than traditional embryo morphology. DESIGN: Prospective clinical study. SETTING: Academic human reproduction laboratory. PATIENT(S): One hundred couples undergoing IVF with conventional insemination or ICSI. INTERVENTION(S): Embryo quality was assessed using both pre-embryo pronuclear morphology (zygote scoring or Z-scoring) at the time of fertilization evaluation and standard day 2 and day 3 embryo morphology (number of blastomeres and grading based on degree of fragmentation and blastomere size). MAIN OUTCOME MEASURE(S): We tested two decision models, one based on Z-scores and another on morphology, to determine which grading system better predicted pregnancy outcomes in assisted reproductive technique. Zygote score and embryo morphology were measured for all embryos and the transferred embryo pool. Implantation and pregnancy rates resulting from the embryo transfers of all cycles were calculated. RESULT(S): The Z-score distribution of 552 embryos was 27% Z1, 8% Z2, 50% Z3, and 15% Z4. Z1 and Z3 embryos had significantly (P approximately .03) higher quality over Z2 and Z4 embryos. Using the Z-score decision model with Z1 embryos having highest priority for transfer, pregnancy rates were similar between Z1 and Z3 embryos. Using embryo morphology as a decision model, pregnancy rates were highest in transfers containing one or two "best"-quality embryos. CONCLUSION(S): Z1 and Z3 embryos had similar morphology and pregnancy rates. The decision model based on the Z-score model was not better than standard embryo morphology in predicting pregnancy outcome.

Adult↗
PubMed1999

Selection of good embryos for transfer depends on embryo cohort size: implications for the 'mild ovarian stimulation' debate.

Embryo quality evaluated by the embryo morphology is a critical parameter in human in-vitro fertilization (IVF) and embryo transfer. It determines which and how many embryos will be replaced, as pregnancy rates are directly related to number and quality of transferred embryos. This retrospective analysis included 1301 IVF and embryo transfer cycles to identify which factors influenced embryo quality. Embryo quality did not correlate with maternal age, causes of infertility, ovarian stimulation parameters or embryo cohort size. However, the mean score of transferred embryos was significantly higher for patients with more than five embryos compared to fewer than five embryos (P < 0.001), irrespective of maternal age. Patients tended to produce a similar embryo quality from cycle to cycle, r = 0.33 (P < 0.001) for the embryo cohort and r= 0.47 (P < 0.001) for the transferred embryos. Poor embryo morphology probably reflects oocytes with compromised development competence and could be an independent factor of infertility. Furthermore, a large embryo cohort was the main factor increasing the chances of at least one good embryo in the cohort.

Adult↗
PubMed1994

Comparative study of the survival rate of frozen-thawed embryos with perforated zonae from microsurgical fertilization and frozen-thawed embryos with intact zonae.

OBJECTIVE: To compare the survival rate of frozen-thawed embryos with perforated zonae from microsurgical fertilization (subzonal insemination) (SUZI) with that of embryos with intact zonae. DESIGN: Thirty-eight embryos resulting from microsurgical fertilization by SUZI were cryopreserved in 16 patient cycles. Within the same period, 140 zonae-intact embryos from 46 patient cycles were cryopreserved. The survival rate of the SUZI embryos was compared with the zonae-intact embryos after thawing. Clinical pregnancies were compared after the transfer of the thawed embryos. RESULTS: The total survival rates were 94.7% and 89.3% for thawed SUZI embryos and zonae-intact embryos, respectively. Within each type of embryo, total survival rates were similar irrespective of the age of the embryos at freezing. The blastomere loss per thawed embryo was the same for zonae-intact and SUZI embryos. One clinical pregnancy was obtained in patients who received thawed SUZI embryos and nine in patients who received embryos with intact zonae. CONCLUSIONS: It is concluded that SUZI embryos can be cryopreserved and thawed with the same degree of confidence as zonae-intact embryos. The low implantation rate of thawed SUZI embryos requires confirmation in a larger clinical series.

Cryopreservation↗
PubMed1988

An assay using embryo aggregation chimeras for the detection of nonlethal changes in X-irradiated mouse preimplantation embryos.

We have developed a short-term in vitro assay for the detection of sublethal effects produced by very low levels of ionizing radiation. The assay utilizes mouse embryo aggregation chimeras consisting of one irradiated embryo paired with an unirradiated embryo whose blastomeres have been labeled with fluorescein isothiocyanate (FITC). X irradiation (from 0.05 to 2 Gy) and chimera construction were performed with four-cell stage embryos, and the chimeras were cultured for 40 h to the morula stage. The morulae were partially dissociated with calcium-free culture medium and viewed under phase contrast and epifluorescence microscopy to obtain total embryo cell number and the cellular contribution of irradiated (unlabeled) and control (FITC labeled) embryos per chimera. In chimeras where neither embryo was irradiated, the ratio of the unlabeled blastomeres to the total number of blastomeres per chimera embryo was 0.50 (17.8 +/- 5.6 cells per unlabeled embryo and 17.4 +/- 5.5 cells per FITC-labeled partner embryo). However, in chimeras formed after the unlabeled embryos were irradiated with as little as 0.05 Gy, the ratio of unlabeled blastomeres to the total number of blastomeres per chimera embryo was 0.43 (P less than 0.01). The apparent decreases in cell proliferation were not observed in irradiated embryos that were merely cocultured with control embryos, regardless of whether the embryos were zona enclosed or zona free. We conclude that very low levels of radiation induce sublethal changes in cleaving embryos that are expressed as a proliferative disadvantage within two cell cycles when irradiated embryos are in direct cell-to-cell contact with unirradiated embryos.

Animals↗
PubMed1997

Embryo morphology or cleavage stage: how to select the best embryos for transfer after in-vitro fertilization.

This retrospective study of 1001 in-vitro fertilization (IVF) cycles included a consecutive series of single transfers (n = 341), dual transfers (n = 410) and triple transfers (n = 250) where all the transferred embryos in each cycle were of identical quality score and identical cleavage stage. In our 2 day culture system, transfer of 4-cell embryos resulted in a significantly higher implantation rate and pregnancy rate (23 and 49%) compared with 2-cell embryos (12 and 22%) and 3-cell embryos (7 and 15%). Furthermore, the transfer of 4-cell embryos resulted in a significantly higher pregnancy rate compared with embryos that had cleaved beyond the 4-cell stage (28%). The implantation rate (21%) and pregnancy rate (43%) after transfer of embryos of score 1.0 were significantly higher than after transfer of embryos of score 2.0 (14 and 32% respectively). Transferring embryos of score 2.1 resulted in significantly higher implantation rates (26%) and similar pregnancy rates compared with score 1.0. Transferring embryos of score 2.2-3.0 resulted in a significantly lower implantation rate (5%) and pregnancy rate (15%). A striking finding was that embryos of quality score 2.0 had a significantly lower implantation rate compared with embryos of quality score 1.0 and 2.1 and a significantly lower pregnancy rate compared to embryos of quality score 1.0. We also found a lower implantation rate and pregnancy rate when transferring 3-cell embryos. These findings may indicate periods of increased sensitivity to damage during the cell cycle. In conclusion, these results substantiate the idea of the superiority of 4-cell embryos and demonstrate that minor amounts of fragments in the embryo may not be of any importance. These findings may call for a shift when weighing the two main morphological components (quality score and cleavage stage) in the sense that reaching a 4-cell cleavage stage even with the presence of a minor amount of fragments should be preferred to a 2-cell embryo with no fragments.

Adult↗
PubMed1999

Secretion of interferon-tau by bovine embryos in long-term culture: comparison of in vivo derived, in vitro produced, nuclear transfer and demi-embryos.

Interferon-tau (IFNtau) is the pregnancy recognition signal of bovine embryos, inhibiting luteolysis. We studied trophoblastic growth and IFNtau secretion of embryos with different developmental potential, i.e., in vivo derived and in vitro produced embryos, cloned embryos and demi-embryos, to evaluate if the ability of secreting IFNtau might be responsible for differences in pregnancy rates after transfer of these categories of embryos to recipients. Day 8 embryos of excellent quality were individually placed in microdrops of buffalo rat liver cell-conditioned medium and maintained for up to 23 days. Embryos were observed on Days 11, 15, 19 and 23, the mean diameter (2r) of attached and spherical embryos was measured, and their trophoblastic area was calculated as r2pi or 4r2pi, respectively. Simultaneously, medium was changed and the IFNtau levels of conditioned media were determined using a bioassay of antiviral activity. Trophoblastic area was smaller (P < 0.05) in demi-embryos than in all other groups, which exhibited similar trophoblastic growth until Day 19. However, on Day 23 trophoblastic area of in vivo derived embryos was more than twice (P < 0.05) as large as those of in vitro produced and nuclear transfer (NT) embryos. IFNtau levels increased only slowly with time in culture of demi-embryos. By contrast, the level of IFNtau doubled from Day 11 to Day 15 in conditioned media from all other groups of embryos. The linear increase in IFNtau production of vivo and in vitro derived embryos continued until the end of the culture period, whereas conditioned media from NT embryos contained significantly (P < 0.05) less IFNtau activity on Days 19 and 23 than those of the former two groups. Our results demonstrate different capabilities of secreting IFNtau for in vivo derived and in vitro produced embryos vs. NT and demi-embryos, which may--at least part--be responsible for the differences in pregnancy rates after transfer to recipients.

Animals↗