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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↗
PubMed2005

Risk of transmission of Mycobacterium avium ssp. paratuberculosis by embryo transfer of in vivo and in vitro fertilized bovine embryos.

Over a 5-year interval, experiments were conducted to determine if Mycobacterium avium ssp. paratuberculosis (Map) is associated with in vivo and in vitro fertilized (IVF) embryos and whether it can be transmitted by embryo transfer. The present studies included: collection of embryos from five asymptomatic, naturally infected donors and transfer to uninfected recipients; collection of oocytes from two naturally infected donors with overt clinical signs; exposure of in vivo and IVF embryos to Map and transfer to uninfected recipients; and the inoculation (transfer) of "clean" IVF embryos to the uterine lumen of infected cows. The presence of Map was confirmed in the uterine horns of all asymptomatic, infected donors. None of the tested embryos, which were not used for embryo transfer, or unfertilized ova (two per batch), were positive for Map, as determined by culture (n = 19) or by PCR (n = 13). However, all in vivo fertilized embryos exposed to Map in vitro (and subsequently sequentially washed) tested positive for Map, by both culture (12 batches) and PCR (15 batches), whereas IVF embryos treated in the same manner tested positive on culture (51%, 18/35 batches) and by PCR (28%, 20/71 batches). Transferring both in vivo embryos and IVF embryos potentially contaminated with Map into 28 recipients resulted in 13 pregnancies and eight calves born without evidence of disease transmission to either the recipients or the offspring over the following 5-year period. In samples collected from one of the clinically infected animals, two of seven (28%) cumulus oocyte complexes (COC) and follicular fluid tested positive by PCR and 10/10 cumulus oocyte complexes on culture for Map. From the second clinically infected cow, three of five batches of IVF embryos (n = 20) were positive on PCR and two of four batches containing unfertilized oocytes and embryos were positive on culture. Only 10% of embryos reached the morula and blastocyst stage 10 days after fertilization. In conclusion, Map is unlikely to be transmitted by embryo transfer when the embryos have been washed as recommended by the International Embryo Transfer Society.

Animals↗
PubMed1983

Effect of the relative locations of embryo and corpus luteum on embryo survival in cattle.

The objectives were to determine if early in gestation (less than day 30) an embryo in the ipsilateral horn increases the survival rate of an embryo in the contralateral horn and if later in gestation (greater than day 30) the presence of two embryos adversely affects the survival of both embryos in cattle. The experiment involved surgical embryo transfer. Sixty Holstein heifers were assigned to 4 groups: group 1) sham bilateral transfers, group 2) one embryo inserted ipsilateral to CL, group 3) one embryo inserted contralateral to CL, and group 4) one embryo inserted into each horn. Diagnosis of pregnancy and ovarian activity were recorded from day 7 or 8 (surgery) to day 110. Results indicated that the rate of embryo survival early in pregnancy (less than day 30) in heifers with the embryo inserted contralateral to the CL (33%, group 3) was increased (P = 0.05) when an embryo was also inserted into the ipsilateral horn (67%, group 4). The hypothesis that the ipsilateral embryo would have a protective effect on the contralateral embryo was, therefore, supported. This may have been due to the prevention of luteolysis by the ipsilateral embryo. However, when an embryo was present in each horn considerable loss occurred greater than day 30 resulting in a low survival rate by day 110 (27%). The hypothesis that the presence of two embryos would decrease the survival probability of both embryos later in gestation was, therefore, supported.

Animals↗
PubMed2004

Uterine position at real embryo transfer compared with mock embryo transfer.

BACKGROUND: The purpose of this study was to determine the consistency in the uterine position between mock and real embryo transfer. METHODS: We reviewed 996 consecutive embryo transfer cycles (585 patients); 74% of patients had an anteverted (AV) uterus and 26% had a retroverted (RV) uterus at mock embryo transfer. All mock and real embryo transfers were performed under abdominal ultrasound guidance. RESULTS: Of 623 fresh embryo transfers in patients with an AV uterus at mock embryo transfer, only 2% became RV, while 55% of 213 embryo transfers in patients with an RV uterus on mock embryo transfer converted to AV at real embryo transfer (P < 0.0001). For frozen-thawed embryo transfer, 12% of AV uteri at mock embryo transfer became RV, while 33% of RV uteri became AV (P = 0.01). CONCLUSIONS: Our data suggest that an RV uterus at mock embryo transfer will often change position at real embryo transfer. Misdirecting the embryo transfer catheter can be avoided by accurate knowledge of the uterine position at the time of embryo transfer, which can be more accurately assessed by routine ultrasound guidance. Additionally, patients with an RV uterus at mock embryo transfer should still present with a full bladder for embryo transfer, since a significant number will convert to an AV position.

Cryopreservation↗
PubMed2001

The impact of the zona pellucida thickness variation of human embryos on pregnancy outcome in relation to suboptimal embryo development. A prospective randomized controlled study.

BACKGROUND: The study represents a prospective randomized controlled study evaluating zona pellucida thickness variation (ZPTV) measurements versus conventional selection of embryos using classic embryo score criteria, prior to embryo transfer in human IVF/intracytoplasmatic sperm injection (ICSI). METHODS: Eighty-six patients having > or =3 embryos, with a classic embryo score of < or =2.2, were allocated to either ZPTV measurement or classic embryo morphology score before embryo transfer. The technician selecting embryos using classic embryo scoring was not aware of the ZPTV measurement results. Of the embryos allocated to ZPTV measurements, only the embryos with the highest ZPTV were transferred. RESULTS: We found no differences in the pregnancy rate per embryo transfer between the two groups (34.4 versus 35.7%). Neither did the implantation rates differ. However, significantly better results were obtained when ZPTV was used as the selection criteria in cases where all embryos had an asynchrony in development or a high embryo score (i.e. were of poorer quality) by classical evaluation (odds ratio = 2.51, confidence interval = 0.33-198). CONCLUSIONS: Using a normally-developed embryo with an optimal embryo score, no beneficial effect of using ZPTV measurement was seen. However, when only less optimal embryos were available to select for transfer, ZPTV provided a x 2.5 increase in the chance of achieving a clinical pregnancy.

Adult↗
PubMed1989

Using the amniotic cavity of the developing chick embryo for the in vivo culture of early-stage mammalian embryos.

The fertile chicken egg may provide an effective, inexpensive method for promoting the development of early-stage embryos from other species. Presently, the loss of viability associated with the in vitro culture of mammalian embryos is hindering the use of in vitro fertilization with farm animals. Consequently, alternative in vitro laboratory methods are needed for the culture of mammalian embryos. A new method has been developed that involves the culture of mammalian embryos in the amniotic cavity of a developing chick embryo. Chick embryos were placed into shell-less incubation (37 C) at the 72-h developmental stage. After 24 h of shell-less incubation, agarose-embedded mammalian embryos were injected into the amniotic cavity of the chick embryo. The mammalian embryos were first placed into a drop of liquid agarose. One to four embryos were then aspirated into a beveled injection pipette and cooled, allowing the agarose to harden. Following penetration of the amnion with the beveled pipette, the agarose cylinder containing the embryos was expelled into the amniotic cavity. The shell-less culture system was then returned to incubation at 37 C for an additional 72 to 96 h. Following incubation, the amniotic cavity containing both chick and mammalian embryos was isolated and the agarose-embedded mammalian embryos were harvested. Significantly more embryos developed in the chick embryo amnion than in the control medium alone. Results obtained using this method on laboratory animals (mice) and on domestic mammals (goats and cattle) indicate that the chick-embryo amnion can support the development of early-stage, mammalian embryos to the blastocyst stage of development.(ABSTRACT TRUNCATED AT 250 WORDS)

Amnion↗
PubMed1996

The influence of number of embryos transferred on pregnancy outcome in women undergoing in vitro fertilization and embryo transfer (IVF-ET).

The purpose of this study was to retrospectively examine the influence of number of embryos transferred on pregnancy outcome in women undergoing in vitro fertilization and embryo transfer (IVF-ET) between July 1990-December 1991 at the Royal Free Hospital, London, United Kingdom. This study included three groups of patients: group 1-the number of embryo used for transferring was one embryo, group 2-the number of embryos used for transferring was two embryos and group 3-the number of embryos used for transferring was three embryos. This study demonstrated that the clinical pregnancy rate increased significantly when transferring two or three embryos compared to one. But this rate did not differ significantly when two embryos were compared with three embryos. The multiple pregnancy rate, miscarriage rate, and ectopic pregnancy rate did not differ significantly among the three groups studied but there was a risk of higher order multiple pregnancy if more embryos were transferred. In this study we found 3 triplets out of 35 pregnancies when 3 embryos were transferred (Group 3). In conclusion, this study demonstrates that when two or three embryos were transferred the clinical pregnancy rate increased significantly. Transferring two embryos instead of three did not alter the pregnancy rate. Moreover, triplets pregnancies were eliminated. Therefore, we suggest limiting the number of embryos to be transferred at two and the supernumerary embryos should be cryopreserved for future transfer.

Adult↗
PubMed2005

Gene expression and developmental competence of bovine embryos produced in vitro under varying embryo density conditions.

The objective of this study was to determine the effects of oocyte/embryo density (embryo number:medium volume ratio) on developmental competence and relative transcription of developmentally important genes in in vitro-produced bovine embryos. Using standard procedures, bovine oocytes were matured, fertilized and cultured in vitro in groups of 5, 10, 20 or 30 oocytes/embryos per 100 microL medium drop (embryo number/medium volume ratios of 1:20, 1:10, 1:5 and 1:3.3, respectively). A semi-quantitative RT-PCR assay was used to determine the relative abundance of Hsp 70.1 and Glut-1 transcripts in Day-6 morulae, and in Day-7 and Day-8 blastocysts and expanded blastocysts. Cleavage rates (55.9, 49.1, 47.1 and 34.6%, respectively) and blastocyst developmental rates (24.1, 18.9, 20.4 and 12.6%, respectively) were higher in groups containing 5, 10 and 20 embryos versus 30 embryos per drop, but there were no significant differences among groups in hatching rates. The relative abundance of Hsp 70.1 transcripts in Day-7 blastocysts was higher in embryos produced in the 20-embryo than in the 5-embryo groups. There were no differences among embryos from all groups in the amount of Glut-1 mRNA (regardless of embryonic stage). However, there were differences in the pattern of Hsp 70.1 gene expression between distinct stages of embryo development and embryo density; these dissimilarities appeared to be associated with compromised development at higher embryo densities. In conclusion, the increase in embryo density during IVM, IVF, and IVC phases of in vitro embryo production might negatively affect embryo development and the pattern of gene expression of in vitro-produced bovine embryos.

Animals↗
PubMed2001

The Graduated Embryo Score (GES) predicts blastocyst formation and pregnancy rate from cleavage-stage embryos.

BACKGROUND: Embryo morphology and cleavage rates alone do not consistently identify embryos with high implantation potential following IVF. Blastocyst transfer has been reported to improve success rates by identifying potentially superior quality embryos. Algorithms for predicting IVF outcomes based on the presence of early developmental milestones have been proposed. Here we introduce the Graduated Embryo Score (GES). METHODS: Nucleolar alignment along the pronuclear axis, regular cleavage and degree of fragmentation at the first cell division, and cell number and morphology on day 3 were weighted to create a possible GES of 100 for each of 1245 fertilized embryos derived from 109 patients aged <40 years. The GES was correlated with IVF outcome. RESULTS: Of 983 embryos for extended culture, 349 (36%) developed to blastocyst and 180 (18%) were good quality (grade I-II). When ranked by cell number and morphology alone, 34% of embryos with > or =7 cells and <20% fragmentation formed good quality blastocysts. Using GES, embryos scoring 90-100 had 64% blastocyst formation compared with 31% scoring 70-85 and with 11% scoring 30-65. Embryos scoring 70-100 had 44% blastocyst development compared with 9% scoring 0-65. Fifty-six patients (51%) conceived on-going gestations from 294 transferred embryos. In patients with at least one transferred embryo scoring > or =70, the pregnancy rate was 59% compared with 34% if all embryos scored <70. The overall implantation rate was 28%. Among embryos scoring 70-100, an implantation rate of 39% was seen, compared with 24% among embryos scoring 0-65. CONCLUSIONS: Predicting which cleaved embryos will form blastocysts could permit the high success rates associated with blastocyst transfer to be achieved from day 3 embryo transfer.

Adult↗
PubMed2005

Comparison of the survival of human biopsied embryos after cryopreservation with four different methods using non-transferable embryos.

BACKGROUND: The standard embryo cryopreservation method is still less than optimal for biopsied embryos. The aim of this study was to compare the survival of biopsied embryos cryopreserved with four different methods using non-transferable embryos. METHODS: Abnormal embryos from one or three pronuclei and spare embryos of grade 3 and 4 were used for this study. Non-biopsied embryos were cryopreserved using the standard method as control. Biopsied embryos were cryopreserved using four methods as follows: standard method, modified freezing method, modified thawing method and vitrification. Blastomere survival and blastulation of frozen-thawed embryos were compared between the different methods. RESULTS: The proportion of embryos with > or = 50% blastomere survival and total blastomere survival rate of biopsied embryos were significantly higher with vitrification than the other three methods. Both the modified freezing and modified thawing methods had significantly higher embryo survival and total blastomere survival rates than standard methods. However, there was no significant difference in blastulation of surviving embryos in all the five groups. CONCLUSIONS: Non-transferable embryos derived from clinical IVF/ICSI are useful for evaluation of the optimal freezing procedures for biopsied embryos. Vitrification increases the survival rate of human biopsied embryos above standard and modified cryopreservation methods.

Biopsy↗
PubMed1981

In vitro studies of mouse embryos bearing mutations in the T complex: effects of culture in suboptimal medium upon t6/t6 and normal embryos.

We have compared the behaviour of normal and t6/t6 embryos in PCMF, a suboptimal 'delay' medium which arrests normal development. Morphologically, the two types of embryos are indistinguishable in this medium. Although beta-glucuronidase activity in embryos does not rise as quickly in delay medium as in cNCTC, a nutritive medium, the rate of increase is similar in t6/t6 and normal embryos. We conclude from these observations that the lethality of t6/t6 embryos is not a consequence of their reaching a given absolute age. Together with previous studies, our data suggest that embryo lethality correlates more closely with metabolic state than with morphological stage. Blastocysts maintained in PCMF are unable to give rise to trophoblast outgrowths but do so upon transfer into cNCTC medium. When a mixture of fourth-day t6/t6 and normal embryos are transferred to cNCTC medium after lengthy pre-incubation periods in PCMF, trophoblast outgrowth is initiated from both types of embryos at approximately the same time. However, when embryos are removed from the genital tract on the second or third day of gestation, t6/t6 embryos are slower to produce trophoblast outgrowths than are normal embryos upon transfer from PCMF to cNCTC medium. Although the reason for this differential behaviour is not yet clear, it is hypothesized that some product(s) required for the outgrowth process is (are) more unstable in third-day t6/t6 embryos than in normal third-day embryos or fourth-day t6/t6 embryos. Our ability to separate t6/t6 from normal embryos by their delayed initiation of trophoblast outgrowth provides us with a convenient way to identify, and to isolate for analysis, enriched populations of homozygous mutant embryos prior to the time at which they show gross morphological abnormalities.

Animals↗
PubMed1997

Comparison of effects of albendazole sulfoxide on in vitro produced bovine embryos and rat embryos.

OBJECTIVE: To evaluate and compare effects of albendazole sulfoxide (ABZSO) on rat embryos and bovine embryos produced in vitro. ANIMALS: In vitro produced bovine embryos. Rat embryos recovered from naturally bred Sprague-Dawley rats. PROCEDURE: 4- and 8-cell bovine embryos were randomly allocated to ABZSO or vehicle control groups. After 48 hours, embryos were evaluated for cell number and blastomere morphology. Rat embryos of similar stages, flushed from the uterine tube on gestational day 2-5, were randomly allocated to treatment or control groups. After 24 hours, embryos were evaluated as described previously. RESULTS: 44% of control bovine embryos divided in culture (> or = 16-cell stage). Fifteen percent of the controls had morphologic abnormalities, including disparity in blastomere size and cytoplasmic vacuoles and stippling. Treated (> or = 1 microgram of ABZSO/ml) bovine embryos differed (P < 0.0001) from controls, with 4% development and 93% abnormal morphology. Forty-five percent of control rat embryos divided in culture. Treated (> or = 500 ng of ABZSO/ml) rat embryos differed (P < 0.0003) from controls with regard to ability to divide. There were no consistent morphologic abnormalities in rat embryos. CONCLUSIONS: In vitro produced bovine embryos were susceptible to ABZSO at a concentration > or = 1 microgram/ ml, resulting in decreased ability to divide and presence of gross morphologic abnormalities. Rat embryos produced in vivo and exposed in vitro to ABZSO at a concentration > or = 500 ng/ml had decreased ability to divide in culture. CLINICAL RELEVANCE: Despite severe effects of ABZSO (> or = 1 microgram/ml) on bovine embryo development in vitro, it is beyond the scope of this study to speculate whether a therapeutic dosage of albendazole (10 mg/kg of body weight) would result in necessary concentrations of ABZSO in vivo to disrupt embryogenesis.

Abnormalities, Drug-Induced↗
PubMed1999

Ultrarapid cryopreservation of human embryos: experience with 1,582 embryos.

OBJECTIVES: To demonstrate the effectiveness of ultrarapid cryopreservation in human embryos. DESIGN: Retrospective study. SETTING: An IVF unit of a medical center. PATIENT(S): All cases received ultrarapid freezing (URF) of embryos and frozen embryo transfer. INTERVENTION(S): Embryos were placed in phosphate-buffered saline (PB1) + 20% maternal serum (MS) for 5 minutes. Embryos were loaded to the straws containing PB1 + MS + 0.25 M sucrose (SUC) + 3.5 M dimethyl sulfoxide for 3 minutes, and then were stored in a liquid nitrogen tank. Embryos were thawed in a 37 degrees C water for 6 seconds and then cultured in PB1 + MS + SUC for 10 minutes. Embryos were transferred into PB1 + MS for 5 minutes and were transferred into patients. MAIN OUTCOME MEASURE(S): The embryo grades before and after URF, the survival and transferred embryo numbers, and the pregnancy and abortion rates were analyzed. RESULT(S): A total of 1,582 embryos were thawed, of which 1,273 (80.5%) embryos were transferred and 1,032 (65.2%) embryos survived with > or = 50% intact blastomere. The embryo numbers with grade I or II before and after freezing and thawing were 1,110 (70.2%) and 790 (50.0%). The mean embryo numbers per transfer was 5.0. Twenty-eight pregnancies (11.4% per transfer) were established, which included 5 miscarriages, 1 ectopic pregnancy, 4 preterm and 18 term deliveries. CONCLUSION(S): Ultrarapid freezing is worthy of reconsideration for embryo cryopreservation.

Abortion, Spontaneous↗
PubMed1997

The presence of a sponsoring embryo in a batch of poor quality thawed embryos significantly increases pregnancy and implantation rate.

OBJECTIVE: To evaluate quantitatively the effect of one good-quality (sponsoring) embryo in a batch of low-quality thawed embryos on the implantation and pregnancy rates (PR). DESIGN: Retrospective analysis of data. SETTING: Tertiary care center IVF clinic affiliated with a university medical school. PATIENT(S): Between March 1988 and April 1995, 392 IVF patients underwent a total of 440 thawing and ET cycles of 1,436 multicellular embryos. MAIN OUTCOME MEASURE(S): Implantation, clinical pregnancy, and multiple pregnancy rates. RESULT(S): In the absence of sponsoring embryos in the thawed batch of embryos, a PR of 9.8% with an implantation rate of 3.1% was achieved. In the presence of a single sponsoring embryo, the PR nearly doubled (18.2%), with a significantly higher implantation rate of 7.0%. Only singleton pregnancies were achieved in the absence of sponsoring embryos compared with 21.7% multiple pregnancies in the single sponsoring embryo group. CONCLUSION(S): The presence of a sponsoring embryo in a batch of poor quality thawed embryos is an important factor that significantly increased pregnancy and implantation rates. The optimal strategy for planning batches of multicellular frozen embryos is to include at least one sponsoring embryo in each batch when possible. We speculate that the sponsoring embryo may favorably influence the chances of low-quality embryos to undergo successful implantation.

Adult↗
PubMed2003

The predictive value of using a combined Z-score and day 3 embryo morphology score in the assessment of embryo survival on day 5.

BACKGROUND: The purpose of this study was to evaluate the ability of using the Z-score alone, or, in combination with the day 3 embryo morphology score, to predict embryo viability at day 5 from a large cohort of embryos derived from patients undergoing treatment with IVF/ICSI. METHODS AND RESULTS: In this retrospective study, a total of 1894 zygotes from 346 treatment cycles (295 couples) was analysed between January 2001 and May 2002. The Z-scoring system was useful in predicting day 5 embryo survival. The mean +/- SD day 5 embryo survival rates were 78.2 +/- 1.7, 49.0 +/- 2.5, 21.4 +/- 3.2 and 11.8 +/- 5.6% for Z-1, Z-2, Z-3 and Z-4 zygotes groups respectively. Embryos derived from Z-1 scores and grade I day 3 embryo scores showed the best day 5 embryo survival and a very high implantation potential. CONCLUSIONS: These data suggest that a combined evaluation of the Z-score and day 3 embryo morphology is highly predictive of embryo outcome after IVF/ICSI. The Z-score could be of great help in the selection of embryos for cultures extended to later stages. The Z-score alone, or preferably in combination with day 3 embryo morphology, is useful in the determination of the most suitable embryos and the number of embryos for transfer, thus achieving the optimal chance of conception while reducing the risk of high order multiple pregnancy.

Adult↗
PubMed1992

Embryo density and medium volume effects on early murine embryo development.

BACKGROUND: One-cell mouse embryos were used to determine the effects of drop size and number of embryos per drop for optimum development in vitro. METHODS: Embryos were collected from immature C57BL6 female mice superovulated with pregnant mare serum gonadotropin and human chorionic gonadotropin and mated by CD1 males. Groups of 1, 5, 10, or 20 embryos were cultured in 5-, 10-, 20-, or 40-microliters drops of CZB under silicon oil at 37.5 degrees C in a humidified atmosphere of 5% CO2 and 95% air. RESULTS: Development score for embryos cultured in 10 microliters was higher than that of embryos cultured in 20 or 40 microliters. Embryos cultured in groups of 5, 10, or 20 had higher development scores than embryos cultured singly. The highest development score was obtained by the combination of 5 embryos per 10-microliters drop. The percentage of live embryos in 20 or 40 microliters was lower than that of embryos cultured in 10 microliters. Additionally, the percentage of live embryos cultured singly was lower than that of embryos cultured in groups. CONCLUSIONS: Our results suggest that a stimulatory interaction occurs among embryos possibly exerted through the secretion of growth factors. This effect can be diluted if the embryos are cultured in large drops or singly.

Animals↗
PubMedDate not supplied

[The development of mouse embryos in vitro in a protein-free medium depending on the numbers of embryos in a microvolume of the medium].

We studied the development of mouse embryos in vitro depending on the number of embryos in a given microvolume of the Ham's F-10 medium without protein or with the addition of serum. The absence of serum from the culture medium did not affect the development of two-cell embryos cultivate in groups of 5-6 (about 90% embryos developed until the stage of blastocyst and over 50% left zona pellucida), but the development of single embryos in the protein-free medium proceeded significantly worse. Single two-cell embryos cultivated in the serum-containing medium developed similar to embryos cultivated in groups. At the same time, no significant differences in development was found between eight-cell embryos cultivated individually or in groups of up to 10 embryos in the Ham's F-10 medium, either in the presence of serum or without it (about 95% embryos developed until the stage of blastocyst and over 70% left zona pellucida). The increase in the number of cultivated embryos over 10 had the adverse effect on development of either two-cell or eight-cell embryos. The attachment of blastocysts to the substrate after leaving zona pellucida and growth of trophectoderm was observed only in the presence of serum. These results suggest that interaction between preimplantation embryos in culture can probably be mediated by factor(s) released by embryos into the medium. Serum appears to contain such factor(s). Sensitivity of embryos to the factor(s) apparently depends on the stage of development.

Animals↗