Repetición de la biopsia ante una falla en la amplificación / Repeat biopsy in case of amplification failure

Resumen OBJETIVO: Analizar los posibles factores asociados con las fallas en la amplificación, los desenlaces de la euploidia y clínicos entre los embriones con repetición de la biopsia y los de una sola (grupo control). MATERIALES Y MÉTODOS: Estudio retrospectivo y multicéntrico de análisis de biopsias de blastocistos practicadas en 22 centros de reproducción asistida (noviembre 2017 a febrero 2022). Se analizaron 4,106 blastocistos procedentes de 1,007 ciclos de ICSI con prueba genética para aneuplidias previa a la implantación. En los blastocistos reportados con falla en la amplificación se analizó el Centro donde se practicó la biopsia, el día en que ésta se tomó, la calidad embrionaria y la incidencia de complicaciones durante el procedimiento. Los resultados se compararon con la prueba genética para aneuploidias previa a la implantación y los desenlaces clínicos entre los embriones con repetición de la biopsia y el grupo control. RESULTADOS: En el 96.0% (3,942) de los embriones se obtuvo resultado y en el 4.0% (n = 164) se reportó falla en la amplificación. La biopsia se repitió en las 99 fallas en la amplificación y se obtuvo resultado en el 83.8% de los casos. Las tasas de euploidia fueron similares entre embriones con repetición de la biopsia y los controles (34.9 en comparación con 39.7%; p > 0.05). El Centro fue el único factor que mostró diferencias en las tasas de falla en la amplificación (p < 0.05). No se observaron diferencias en el día de la biopsia o la calidad embrionaria. Las tasas de embarazo (51.0 en comparación con 58.3%), implantación (63.9 en comparación con 61.5%) y aborto (16.9 en comparación con 28.6%) fueron similares entre embriones con una sola biopsia o repetición de ésta, respectivamente. CONCLUSIONES: El Centro fue el principal factor que influyó en las fallas en la amplificación. Las tasas de euploidia y los desenlaces clínicos no difirieron entre el grupo control y los embriones con repetición de la biopsia; por consiguiente, se recomienda repetir la biopsia en los embriones con falla en la amplificación.

Abstract OBJECTIVE: To analyze possible factors associated with amplification failures, euploidy and clinical outcomes between repeat and single biopsy embryos (control group). MATERIALS AND METHODS: Retrospective multicenter study involving 4,106 blastocysts from 1,007 ICSI cycles with preimplantation genetic testing for aneuploidy performed by next generation sequencing. In case of DNA amplification failure, the IVF center where biopsies were performed, the day of biopsy, the embryo quality and the incidence of complications during biopsy were analyzed. Preimplantation genetic testing for aneuploidy results and clinical outcomes were compared between re-biopsied embryos and the control group. RESULTS: Of the 4,106 blastocysts included in this study, 96.0% (3,942) obtained a result while 4.0% (164) had an amplification failure. Ninety-nine embryos with amplification failure were re-biopsied and 83.8% resulted in an informative diagnosis. Euploidy rates were equivalent between re-biopsied and control blastocysts (34.9% vs 39.7%, P>0.05). The only factor significantly affecting the amplification failure rates was the IVF center. No differences were observed between biopsy days or embryo quality. Pregnancy (51.0% vs 58.3%), implantation (63.9% vs 61.5%) and miscarriage rates (16.9% vs 28.6%) were similar between single and repeat biopsied embryos, respectively. CONCLUSIONS: The centre was the main factor influencing amplification failures. Euploidy rates and clinical outcomes did not differ between the control group and repeat biopsied embryos; therefore, repeat biopsy is recommended for embryos with amplification failure.

Effect of Insemination Methods on Pregnancy Outcomes of Day 6 Blastocyst Transfer in Frozen Cycles / 中山大学学报(医学科学版)

ObjectiveTo compare the effects of two different insemination methods， conventional in vitro fertilization （IVF） and intracytoplasmic sperm injection （ICSI）， on pregnancy outcomes in patients with frozen-thawed D6（day 6） blastocyst transfer. MethodsA retrospective cohort study was conducted to analyze the clinical data of patients with thawed D6 blastocyst transfer between January 2018 and April 2020 at the Fertility Center of the Third Hospital of Sun Yat-sen University， divided into conventional IVF group （446 cycles ） and ICSI fertilization group （200 cycles） according to the fertilization method. Patients were divided into those with a history of D5（day 5） blastocyst transfer and those without. The patients’ general characteristics， blastocyst quality， and pregnancy outcomes of the two groups were compared. ResultsBMI， years of infertility， and basal FSH were not statistically significant in the IVF and ICSI groups （P > 0.05）. Regardless of the history of D5 transfer， patients in the ICSI group were younger than those in the IVF group （P < 0.001）， the proportion of primary infertility was significantly higher in the ICSI group （P < 0.001）， and the number of oocytes obtained and the number of normally fertilized oocytes in the ICSI group were higher than those in the conventional IVF fertilization group （P < 0.001）. The proportion of stage V and Ⅵ blastocysts was significantly higher in the conventional IVF group than in the ICSI group （21.6 % vs. 3.14 %， P < 0.001）. High-quality blastocysts with an ICM score of A were significantly higher in the ICSI group than in the IVF group （23.8 % vs. 14.3 %， P = 0.01）. The HCG-positive and clinical pregnancy rates were significantly higher in the ICSI group than in the IVF group （65.5 % vs. 48.4 %， P < 0.001； 56 % vs. 41.3 %， P = 0.001）， and embryo implantation and live birth rates were also higher in the ICSI group than in the conventional IVF group （43.8 % vs. 30.9 %， P < 0.001； 43.0 % vs. 31.8 %， P = 0.006）. After correcting for age and number of oocytes obtained between the two groups， the clinical pregnancy rate was still significantly higher in the ICSI group than in the conventional IVF group （OR： 1.590， 95 % CI： 1.030， 2.455， P = 0.036）. Infant birth weight was lower in the ICSI group than in the IVF group （P = 0.016）， and the differences in preterm birth rate， sex ratio， and mode of delivery were not statistically significant between the two groups. ConclusionsClinical pregnancy and live birth rates after thawing and transfer of D6 blastocysts fertilized by ICSI are higher than those of D6 blastocysts fertilized by conventional IVF， which may be related to the different factors contributing to the slow development of blastocysts in patients who received different fertilization methods. The relatively good pregnancy outcome after the transfer of thawed D6 blastocysts fertilized by ICSI may compensate to some extent for the difference in pregnancy outcome due to the relatively slow blastocyst development and a relatively higher proportion of D6 blastocysts after ICSI fertilization in male infertility patients.

Correlation Between Blastocyst Morphology Score， Early Serum β-hCG Concentrations and Live Birth After Thawed Single Blastocyst Transfer / 中山大学学报(医学科学版)

ObjectivesTo assess the correlation between blastocyst morphology score， serum human chorionic gonadotropin β subunit （β-hCG） levels on day 12 after transfer and live birth outcomes among cycles tested HCG-positive after thawed single blastocyst transfer； to analyze the predictive value of serum β-hCG levels on live birth. MethodsWe reviewed the data of 519 frozen-thawed single blastocyst transfer cycles （FET） that had been tested HCG-positive from January 2016 to May 2020 at our IVF center. These FET cycles were firstly divided into 4 groups （AA， AB， BA， and BB） according to Gardner's grading system of inner cell mass （ICM） and trophectoderm cell （TE）， and then 4 groups （stages 3， 4， 5 and 6） according to the degree of blastocyst expansion. Serum β-hCG concentrations on day 12 after transfer and live birth rates were compared among groups transferred with different blastocysts grading and expansion stage. The relationship between Gardner’s grading or expansion stage of blastocysts and serum β-hCG levels was determined by correlation test， and ROC curves were plotted to determine the threshold values of serum β-hCG for predicting live birth. Results（1） The serum β-hCG concentration in the AA group and AB group on the 12th day after the transfer was significantly higher than that in the BB group （P <0.001， P <0.001）. However， there was no significant difference in the live birth rate when different ICM/TE-graded blastocysts were transferred （P = 0.120）. There were no significant differences in serum β-hCG concentration on day 12 after transfer and live birth rate among blastocysts with different expansion stages （P = 0.091， P = 0.557）. （2） There was a significant weak correlation between blastocyst ICM/TE grading and serum β-hCG concentration on day 12 （rs = -0.221， P <0.001）， and even after controlling for confounding factors （ rs = -0.228， P <0.001）；There was no significant correlation between blastocyst’s expansion stage and serum β-hCG concentration on day 12 after the transfer （rs = -0.052， P = 0.240）， and the association remained insignificant after controlling for confounding factors （rs = -0.029， P = 0.508）. （3） ROC curve analysis showed that the cut-off value for predicting live birth by serum β-hCG on day 12 was 657.5 mU/mL （P < 0.001）. ConclusionsNeither the ICM/TE grade nor the expansion stage of blastocysts affect the live birth rate，there is significant difference in the level of β-hCG produced by blastocyst with different ICM/TE grade；Our results suggest that early serum β-hCG level can predict live birth.

Adopción embrionaria: estado de la cuestión ético-jurídica en el Perú

La técnica de reproducción asistida trae consigo un número de embriones sobrantes. Se propone una revisión del estado de la cuestión de estos embriones en el Perú. Se revisa desde las perspectivas biológica, ética y jurídica. Se profundiza en el ordenamiento jurídico peruano, sobre el estado de desprotección de estos embriones y se contrasta con una posible solución: la adopción embrionaria. Se constata el estado de incertidumbre de la situación de estos embriones y se concluye como ilegítima la práctica de la adopción embrionaria, por vulnerar la dignidad y condición de sujeto de derecho del embrión sobrante.

The assisted reproduction technique brings with it several surplus embryos. A review of the situation of these embryos in Peru is proposed from biological, ethical, and legal perspectives. We delve into the Peruvian legal system regarding the lack of protection of these embryos and contrast it with a possible solution: embryonic adoption. The uncertainty of the situation of these embryos is verified, concluding that the practice of embryonic adoption is illegitimate, for it constitutes a violation of the dignity and rights-holder condition of the remaining embryos.

A técnica de reprodução assistida traz consigo uma série de embriões excedentes. Propõe-se uma revisão da situação desses embriões no Peru. É feita uma revisão a partir de perspectivas biológicas, éticas e legais. Aprofunda-se no ordenamento jurídico peruano sobre o estado de desproteção desses embriões e contrasta-se com uma possível solução: a adoção embrionária. Verifica-se o estado de incerteza da situação desses embriões e conclui-se como ilegítima a prática da adoção de embriões, por violar a dignidade e condição de sujeito de direito do embrião remanescente.

Derechos del concebido

La fecundación consiste en la fusión de los gametos masculino y femenino en el tercio externo de la trompa uterina para constituir el cigoto, que inicia su división celular, viaja al útero, se implanta como blastocito en el endometrio a los siete días y comienzan los procesos de embriogénesis y morfogénesis. Concepción es la acción o efecto de quedar embarazada una mujer. En el tema de los derechos establecidos al concebido, el inicio de la vida humana plantea amplia discusión entre las definiciones que plantea la ciencia frente a las de grupos de la sociedad civil. No siendo el cigoto una persona humana, de acuerdo con la Corte Interamericana de Derechos Humanos (CIDH) y otros no sería sujeto de los derechos establecidos para la persona, los cuales estarían íntimamente relacionados a los derechos de la mujer embarazada. La concepción ocurriría cuando el embrión se implanta en el útero y no aplicaría el artículo 4 de la CIDH. La doctrina, la legislación y la jurisprudencia peruana establecen amplia tutela jurídica a favor del concebido e instauran el inicio de la vida a partir de la fecundación. La Sentencia de la CIDH para el caso Artavia Murillo contra Costa Rica introduce la figura española del preembrión, al cual no se le otorga tutela jurídica hasta los 14 días, momento en que la ciencia médica establece la implantación del embrión en el endometrio materno. Para consideraciones éticas actuales, el embarazo humano empieza con la implantación del blastocisto en el endometrio y no existe sustento para aceptar el derecho del concebido desde el momento de la fecundación como si fuera una persona. Empero se señala que la vida humana posee una continuidad ininterrumpida desde su concepción hasta su fin natural, la muerte. El cigoto es vida que se inicia con dotación genética propia diferente a la de sus progenitores. El embrión preimplantatorio solo podrá originar un ser humano que se gesta y desarrolla dentro del cuerpo de otra persona, con propia dinámica vital.

Fertilization consists of the fusion of the male and female gametes in the outer third of the uterine tube to form the zygote, which begins its cell division, travels to the uterus, implants as a blastocyst in the endometrium after seven days and begins the processes of embryogenesis and morphogenesis. Conception is the action or effect of a woman becoming pregnant. About the rights established for the conceived, the beginning of human life raises a wide discussion between the definitions proposed by science and those of civil society groups. Not being the zygote a human person, according to the Inter-American Court of Human Rights (IACHR) and others, it would not be subject to the rights established for the person, which would be closely related to the rights of the pregnant woman. Conception would occur when the embryo implants in the uterus and Article 4 of the IACHR would not apply. Peruvian doctrine, legislation and jurisprudence establish broad legal protection in favor of the conceived and indicate the beginning of life from the moment of fertilization. The Judgment of the IACHR in the case of Artavia Murillo v. Costa Rica introduces the Spanish figure of the pre-embryo, which is not granted legal protection until 14 days, when medical science establishes the implantation of the embryo in the maternal endometrium. For current ethical considerations, human pregnancy begins with the implantation of the blastocyst in the endometrium and there is no basis for accepting the right of the conceived from the moment of fertilization as if it were a person. However, it is pointed out that human life has an uninterrupted continuity from conception to its natural end, death. The zygote is life that begins with its own genetic endowment different from that of its progenitors. The preimplantation embryo can only give rise to a human being that is gestated and develops within the body of another person, with its own vital dynamics.

Chromosomal Abnormalities in Embryos

From patients with a poor prognosis of pregnancy, 1620 embryos generated in vitro and chromosomal analysis was performed on these embryos. The result was yielded in 1596 embryos, out of them 536(34%) were euploid and 1060(66%) carried chromosomal abnormalities. In addition, 92% of embryos with multinucleated cells were diagnosed mosaics whereas the 86% of chromosomal abnormalities were associated to the presence of cytoplasmic concentration. For the derivation of the normal embryonic stem cell (ESC)lines and developmental modelling aneuploid embryos have been used. Genetic diagnosis at the cleavage or blastocyst stage could be partly abnormal because during the preimplantation diploid- aneuploid mosaic embryos was most frequently observed. From a single cell of a particular embryo the chromosomal status of that embryo can be determined, thus the prevalence of mosaicism. Detection of aneuploidy in single cells have been developed recently. After conducting research methods, it was confirmed that aneuploidy is a common feature of human oocytes and preimplantation embryos. The detection of segmental aneuploidy is currently considered problematic for embryo diagnosis and patient counselling, so the data are of great relevance for preimplantation genetic testing. The first major milestone in early mammalian embryogenesis was the formation of a totipotence blastocyst which is capable of implantation. The whole chromosomal abnormalities, or aneuploidy, determines whether the human embryos will arrest or reach the blastocyst stage. Certain embryos can still form blastocyst depending on the type of chromosomal abnormalities and that can be morphologically indistinguishable from chromosomally normal embryos.

Analysis of pregnancy outcomes of single top-quality blastocyst transfer developed from cleavage embryos with different grading / 中华妇产科杂志

Objective:To explore the clinical outcomes of top-quality blastocysts transfer developed from cleavage embryos with different grading and determine whether the cleavage stage embryo morphology grading should be taken into consideration when transferring the embryo at the blastocyst stage.Methods:A number of 3 059 cycles were included with single top-quality blastocyst transfer dating from January 2017 to May 2021 in Henan Provincial People′s Hospital. According to the number of cleavage sphere and degree of fragmentation, all cleavage stage embryos were divided into three groups: top D3 embryo (8 cells, ≤5% fragments)-TB group, suboptimal D3 embryo (8 cells, 5%<fragments≤10%; 7 cells or 9 cells, ≤10%)-TB group, and normal D3 embryo-TB group. Univariate analysis, multivariate logistic regression analysis and threshold effect analysis were performed on the data.Results:The clinical pregnancy rates of top D3 embryo-TB group(1 326 cycles), suboptimal D3 embryo-TB group (830 cycles) and normal D3 embryo-TB group (903 cycles) were 69.53%, 70.12% and 66.67%, respectively ( P>0.05); and the early abortion rate were 10.74%, 12.54% and 12.62%, respectively ( P>0.05). After adjusting for confounders, logistic regression showed that no significant associations were found between cleavage stage embryo morphology grading and clinical pregnancy rate (suboptimal D3 embryo-TB group: OR=1.02, 95% CI: 0.76-1.38, P=0.879; normal D3 embryo-TB group: OR=0.84, 95% CI: 0.61-1.14, P=0.262) and early abortion rate (suboptimal D3 embryo-TB group: OR=1.18, 95% CI: 0.77-1.82, P=0.445; normal D3 embryo-TB group: OR=1.26, 95% CI: 0.81-1.98, P=0.309). The results of threshold effect analysis showed that when a single top-quality blastocysts was transferred, the effect of age on the clinical pregnancy rate showed a curve relationship, when the age was≥33 years old, the clinical pregnancy rate decreased significantly with age increased ( OR=0.89, 95% CI: 0.83-0.95, P=0.007); and there was no significant change in early abortion rate ( OR=1.01, 95% CI: 0.97-1.06, P=0.628). Conclusions:Cleavage stage embryo grading is not found to correlate with clinical outcomes in single top-quality blastcyst tranfer. Therefore, when considering blastocyst transfer, its morphology at blastocyst stage is more relevant. The effect of age on pregnancy outcomes of single blastocyst transfer should be considered.

First Baby Born in Brazil after Simultaneous Diagnosis through Non-Invasive and Conventional PGT-A / Primeiro bebê nascido no Brasil após diagnóstico simultâneo por PGT-A não-invasivo e convencional

Abstract Non-invasive preimplantation genetic testing for aneuploidies (niPGT-A) aiming to assess cell-free embryonic DNA in spent culturemedia is promising, especially because it might overcome the diminished rates of implantation caused by the inadequate performance of trophectoderm (TE) biopsy. Our center is part of the largest study to date assessing the concordance between conventional PGT-A and niPGT-A, and we report here the delivery of the first baby born in Brazil using niPGT-A. The parents of the baby were admitted to our center in 2018. They did not present history of infertility, and they were interested in using in vitro fertilization (IVF) and PGT-A in order to avoid congenital anomalies in the offspring. A total of 11 (3 day-5 and 8 day-6) expanded blastocysts were biopsied, and the spent culture media (culture from day-4 to day-6) from 8 day-6 blastocysts were collected for niPGT-A. Overall, 7 embryos yielded informative results for trophectoderm (TE) and media samples. Among the embryos with informative results, 5 presented concordant diagnosis between conventional PGTA and niPGT-A, and 2 presented discordant diagnosis (1 false-positive and one falsenegative). The Blastocyst 4, diagnosed as 46, XY by both niPGT-A and conventional PGTA, was warmed up and transferred, resulting in the birth of a healthy 3.8 kg boy in February 2020. Based on our results and the recent literature, we believe that the safest current application of niPGT-A would be as a method of embryo selection for patients without an indication for conventional PGT-A. The approximate 80% of reliability of niPGT-A in the diagnosis of ploidy is superior to predictions provided by other noninvasive approaches like morphology and morphokinetics selection.

Resumo Abordagens para o teste genético pré-implantacional não-invasivo para aneuploidias (non-invasive preimplantation genetic testing for aneuploidies, niPGT-A, em inglês) com o objetivo de avaliar o DNA embrionário livre são promissoras, especialmente porque estas podem reverter as menores taxas de implantação causadas por inadequada biópsia de trofectoderma (TE). Nesse contexto, nosso centro é parte do maior estudo atual que avalia as taxas de concordância entre PGT-A convencional e niPGT-A, e relatamos aqui o nascimento do primeiro bebê brasileiro após niPGT-A. Os pais do bebê foram admitidos no nosso centro em 2018. Eles não apresentavam histórico de infertilidade, e estavam interessados em utilizar os tratamentos de fertilização in vitro (FIV) e PGT-A para evitar anomalias congênitas na progênie.Umtotal de 11 blastocistos expandidos (3 do dia-5 e 8 do dia-6) foram submetidos a biópsia, e os meios de cultivo condicionados (cultivo do dia-4 ao dia-6) de 8 blastocistos do dia-6 foram coletados para niPGT-A. No total, resultados informativos para as amostras de TE e dos meios foram obtidos para sete embriões. Entre os embriões com resultado informativo, 5 apresentaram diagnóstico concordante entre PGT-A convencional e niPGT-A, e 2 apresentaram diagnóstico discordante (1 falso positivo e 1 falso negativo). O Blastocisto 4, diagnosticado como 46, XY por ambos niPGT-A e PGT-A convencional, foi desvitrificado e transferido, o que resultou no nascimento de ummenino saudável, que pesava 3,8 kg, em fevereiro de 2020. Com base em nossos resultados e literatura contemporânea, acreditamos que a aplicação atualmais segura do niPGT-A seria como método de seleção embrionária para pacientes sem indicação ao PGT-A convencional. A confiabilidade aproximada de 80% do niPGT-A para determinação da ploidia ainda é superior àquela obtida com abordagens não invasivas, como seleção morfológica ou morfocinética.

Impact of Plasmatic Progesterone on the Day of Frozen Embryo Transfer in Hormone-induced Cycles / Impacto da progesterona plasmática no dia da transferência de embriões congelados em ciclos induzidos por hormônios

Abstract Objective To establish a relationship between serum progesterone values on the day of frozen blastocyst transfer in hormone-replaced cycles with the probability of pregnancy, miscarriage or delivery. Methods This was an ambispective observational study including all frozen-thawed embryo transfer cycles performed at our department following in vitro fecundation from May 2018 to June 2019. The outcomes evaluated were β human chorionic gonadotropin (β-hCG)-positive pregnancy and delivery. Groups were compared according to the level of serum progesterone on the day of embryo transfer: the 1st quartile of progesterone was compared against the other quartiles and then the 2nd and 3rd quartiles against the 4th quartile. Results A total of 140 transfers were included in the analysis: 87 with β-HCG>10 IU/L (62%), of which 50 (36%) delivered and 37 had a miscarriage (42%).Women with lower progesterone levels (< 10.7ng/mL) had a trend toward higher β-HCG-positive (72 versus 59%; p>0.05), lower delivery (26 versus 39%; p>0.05) and higher miscarriage rates (64 versus 33%; p<0.01). Comparing the middle quartiles (P25-50) with those above percentiles 75, the rate of pregnancy was similar (60 versus 57%; p>0.05), although there was a trend toward a higher number of deliveries (43 versus 31%; p>0.05) and a lower number of miscarriages (28 versus 45%; p>0.05). These differences were not statistically significant. Conclusion There were no differences in pregnancy and delivery rates related with the progesterone level when measured in the transfer day. The miscarriage rate was higher in the 1st quartile group.

Resumo Objetivo Avaliar se existe alguma relação entre os valores plasmáticos de progesterona no dia da transferência de um blastocisto desvitrificado em ciclos hormonalmente substituídos e a taxa de gravidez, aborto ou nascido vivo. Métodos Estudo observacional, ambispectivo, incluindo todos os ciclos de transferência de blastocistos congelados no nosso departamento, entre maio de 2018 e junho de 2019. Avaliou-se a taxa de gravidez e de nascidos vivos após 24 semanas de gestação. Os grupos foram comparados de acordo com os valores de progesterona plasmáticos dosados no dia da transferência do blastocisto: comparou-se o 1° quartil com os outros e depois os 2° e 3° quartis com o 4°. Resultados Avaliaram-se 140 transferências: 87 com β gonadotrofina coriônica humana (β-HCG)>10 IU/L (62%), 50 das quais terminaram em nascido vivo (36% do total), enquanto 37 tiveram um aborto (42% das gravidezes). Verificou-se uma tendência para menor número de recém-nascidos nas transferências com níveis de progesterona no 1° quartil (<10.7ng/mL) (26 versus 39%; p>0.05) e ummaior número de abortos (64 versus 33%; p<0.01). Comparando o 2° e 3° quartis com o 4°, verificouse que nos casos em que a progesterona estava acima do percentil 75, apesar de uma taxa de gravidez semelhante (60 versus 57%; p>0.05), houve uma tendência para uma maior taxa de nascidos vivos (43 versus 31%; p>0.05) emenor número de abortos (28 versus 45%; p>0.05) abaixo do percentil 75. Estas diferenças não foram estatisticamente significativas. Conclusão Não se verificaram diferenças estatisticamente significativas para taxa de gravidez e de nascido vivo. A taxa de aborto foi maior no primeiro quartil.

The effect of melatonin on in vitro maturation fertilization and early embryo development of mouse oocytes and expression of HMGB1 gene in blastocysts

Antioxidants are commonly used for maturation, fertilization and early development of embryos. Melatonin as an antioxidant have been recently proven to be useful for the assisted reproductive technology. In the present study, we evaluated the roles of melatonin in the in vitro maturation, fertilization, development and also the gene expression of high mobility group box-1 (HMGB1) in the blastocysts. The immature oocytes of BDF1 mice were transferred to the media containing different doses of melatonin (10-6, 10-9, 10-12 M). The blastocysts that developed under in vitro fertilization from each group were stained to determine the cell number of embryos and analyzed to determine the expression level of HMGB1 by real-time PCR. The most effective doses of melatonin for maturation of oocytes were 10-6 and 10-12M (P<0.05). Fertilization rate, early development and the cell number of blastocysts were significantly higher in the group that treated with 10-12 M of melatonin comparing to the other groups. The HMGB1 expression decreased in groups that treated with 10-6M and 10-9M of melatonin and increased in the group that treated with 10-12 M of melatonin, but did not show a significant difference (p˃0.05). From the results, it may be concluded that the melatonin could be effective when the embryos undergo maturation, fertilization and early developmental processes. The HMGB1 expression, as a marker of early development in mice embryos, increased in the groups that treated with low doses of melatonin

The strategy of frozen-thawed blastocyst transfer in women over 40 years / 中国医师杂志

Objective:To explore the selection strategy of blastocyst transfer number in freeze-thaw cycle for women over 40 years old, so as to provide reference for reducing twin rate and improving perinatal clinical outcome.Methods:A retrospective analysis was made of 377 patients who underwent freeze-thaw blastocyst transplantation in the reproductive center of Guangdong Maternal and Child Health Hospital from January 2017 to December 2019. They were divided into single blastocyst and double blastocyst transplantatio groups according to the number of blastocyst transplantation. The clinical pregnancy rate, implantation rate, abortion rate, live birth rate, premature delivery rate, twin rate and singleton delivery rate were compared between the two groups.Results:⑴There was no significant difference between two groups regarding the majority of baseline characteristics, including age at retrieval, age at transfer, body mass index (BMI), antral follicle count (AFC), basal follicle stimulating hormone (FSH), anti Mullerian hormone (AMH), endometrial thickness at transfer day, number of oocytes retrieved, Gn starting dose, Gn days, Gn dosage, embryos at cleavage stage and top-quality embryos ( P>0.05). ⑵ There was no significant difference in the rate of implantation, early pregnancy loss, late pregnancy loss and live birth between two groups ( P>0.05). ⑶ The preterm birth rate was higher in the double blastocyst transplantation group compared with the single blastocyst transplantation group, albeit not reaching significant difference (31.7% vs 12.5%, P=0.083). ⑷ The clinical pregnancy rate and the twin pregnancy rate was significantly higher in the double blastocyst transplantation group compared with the single blastocyst transplantation group ( P<0.05). ⑸ The singleton birth rate was significantly lower in the double blastocyst transplantation group compared with the single blastocyst transplantation group (75.61% vs 95.83%, P<0.05). Conclusions:In women ≥40 years old, transferring a single blastocyst can result in live birth rate that is similar as transferring two blastocysts while dramatically reducing the risk of twin pregnancy rate and increasing singleton birth rate.

Embryonic Toxicity Test of Medical Devices for Human Assisted Reproductive Technology / 中国医疗器械杂志

OBJECTIVE@#To introduce the test methods of embryo toxicity applied to medical devices for human assisted reproductive technology (ARTMD), and provide the evaluation reference.@*METHODS@#The embryo toxicity test methods of ARTMD were summarized, and the key procedures and challenges in their safety evaluation were also discussed.@*RESULTS@#Establishing sensitive and stable test system is important to guarantee the safety and efficacy of ARTMD.@*CONCLUSIONS@#It remains development opportunities in improving sample preparation, extending test technology and expending evaluation method.

Comparison of fresh and frozen thawed embryo transfer in terms of clinical pregnancy rate

Background: In a standard IVF (in-vivo fertilization) procedure, the embryos formed after the fertilization of male and female gametes are allowed to grow for 3-5 days and then transferred back to the uterine cavity of the female, where they might get attached and start to grow. Objective of this study was to compare clinical pregnancy rate of fresh embryo transfers and frozen-thawed embryo transfers.Methods: This is a retrospective case control study in patients undergoing IVF /ICSI cycles from January 2018 to December 2018 were enrolled in assisted reproduction. Total of 200 women which contains 118 fresh embryo transfers and 82 frozen-thawed embryo transfers are studied.Results: Clinical pregnancy rates of fresh cleavage-stage embryo transfers compared with frozen-thawed cleavage-stage embryo transfers, were (53.3% versus 39.6%). Ectopic pregnancy is also significant in comparison. In patients under 35 years of ages and (57.1% versus 12.5%). In patients older than 35 years old, respectively. The multiple pregnancy rates, abortion rates and ectopic pregnancy rates did not differ significantly among the groups. Multiple pregnancy rate and abortion rate is significantly high in frozen-thawed blastocyst transfer than fresh embryo transfer. Whereas the ectopic pregnancy rates had no difference in both groups.Conclusions: The clinical pregnancy rates in fresh embryo transfer is high than that of frozen-thawed blastocyst.

Evaluación de la tasa de blastocistos euploides en fase folicular y lútea (DuoStim) en mujeres con pobre respuesta ovárica: estudio preliminar en México / Euploid blastocyst rate in follicular vs luteal phase (DuoStim) in women with poor ovarian response: Preliminary study in Mexico

Resumen: OBJETIVO: Comparar la tasa de blastocistos euploides obtenida después de la estimulación ovárica en fase folicular con la fase lútea en un mismo ciclo menstrual en pacientes con deficiente respuesta ovárica. MATERIALES Y MÉTODOS: Estudio clínico, prospectivo y comparativo llevado a cabo en el Centro de Reproducción Arcos, Nascere, entre los meses de enero a julio de 2019. Se incluyeron pacientes con pobre respuesta ovárica según los criterios de Bologna y con indicación de PGT-A. Las estimulaciones en fase folicular y lútea se efectuaron con antagonista de la GnRH y FSHr/LHr (2:1) a partir del día 3 del ciclo y 5 días después de la primera recuperación de los ovocitos. Para completar el proceso de maduración ovocitaria se utilizaron análogos de GnRH, se tomó una biopsia de trofoectodermo en día 5-7. RESULTADOS: Se estudiaron 20 pacientes. Al comparar la fase folicular con la lútea la tasa de fertilización fue de 79% (IC95%: 29-46) vs 55% (IC95%: 34-53), la tasa de blastocistos 42% (IC95%: 19-44) vs 45% (IC95%: 24-55) y la tasa de blastocistos euploides 100% (IC95%: 44-53) vs 70% (IC95%: 38-46), respectivamente. Solo la tasa de recuperación de ovocitos en metafase II mostró diferencias significativas entre ambas fases 40% (IC95%: 18-37) vs 59% (IC95%: 31-59), p = 0.0333 en la fase folicular y lútea, respectivamente. CONCLUSIONES: La estimulación ovárica bifásica (folicular-lútea), en el mismo ciclo menstrual (DuoStim), resultó en mayor tasa de recuperación de ovocitos en metafase II durante la fase lútea. Sin embargo, las tasas de desarrollo embrionario a día 5-6 (blastocistos) y de embriones euploides fueron similares entre ambas fases.

Abstract: OBJECTIVE: Euploid blastocyst rate comparison between ovarian stimulation in follicular vs luteal phase performed in the same menstrual cycle in patients with poor ovarian response. MATERIALS AND METHODS: Clinical, prospective and comparative study conducted at Centro de Reproducción Arcos S.C., "Nascere", during january-july, 2019. Patients with PGT-A indication and poor ovarian response according to Bologna criteria were included. Under a short GnRH-antagonist protocol, stimulations, both in follicular and luteal phase were performed using rFSH/rLH (2:1) from day 3 of the cycle and 5 days after the first oocyte retrieval. In addition, ovulation trigger with an GnRH agonist was used, finally, on day 5-6 of embryo development, trophoctoctoderm biopsy was performed. RESULTS: In this study, 20 patients were included; when comparing follicular phase vs luteal phase, we found that fertilization rate was 79% (95%CI 29-46) vs 55% (95%CI 34-53), blastocysts rate was 42% (95%CI 19-44) vs 45% (95%CI 24-55) and euploid embryo rate was 100% (95%CI 44-53) vs 70% (95%CI 38-46). Only the oocyte recovery rate in metaphase II showed significant differences between both phases 40% (IC 95% 18-37) vs 59% (IC 95% 31-59), p=0.0333. CONCLUSION: Biphasic ovarian stimulation (follicular/ luteal) in the same menstrual cycle (DuoStim) resulted in a higher metaphase II ooctye recovery rate during the luteal phase in comparison with the follicular phase. However, the rates of blastocysts and euploid blastocysts were similar between both phases.

Diagnóstico genético preimplantacional no invasivo; datos preliminares en México / Non-invasive preimplantation genetic diagnosis; preliminary data in Mexico

Resumen: OBJETIVO: Analizar las tasas de concordancia, falsos positivos y negativos entre el ADN embrionario circulante en medio de cultivo y su relación con los reportes de la biopsia de trofoectodermo. MATERIALES Y MÉTODOS: Estudio observacional, prospectivo y comparativo, llevado a cabo en el Centro de Reproducción Arcos Nascere en noviembre 2018. Criterios: de inclusión: parejas en esquema de fertilización in vitro, con diagnóstico genético preimplantacional de aneuploidias. Criterios de exclusión: pacientes con anomalías estructurales o enfermedades monogénicas. Criterio de eliminación: blastocistos con eclosión asistida. Variables de respuesta: tasa de concordancia, falsos positivos y negativos entre las biopsias de trofoectodermo y los medios de cultivo. El análisis estadístico se realizó con SPSS 25.0, con pruebas t de Student y χ2 con valor de p < 0.05 significativa. RESULTADOS: Se analizaron 20 blastocistos de 5 parejas y se obtuvieron resultados informativos de 17 (amplificación global exitosa); 70% en día 5 y 100% en día 6. La tasa general de concordancia entre las biopsias de trofoectodermo y los medios de cultivo fue de 68.7% (42.8% en día 5 y 88.8% en día 6). En cuanto a las discrepancias, solo se observaron 2 falsos negativos en los medios de cultivo vs la biopsia de trofoectodermo (14.2% en día 5 y 11.11% en día 6); hubo 3 casos de falsos positivos (la mitad en día 5 y ninguno en día 6-7). CONCLUSIONES: Con la prueba genética no invasiva de aneuploidias se alcanzaron altas tasas de concordancia, sobre todo en embriones en día 6.

Abstract: OBJECTIVE: Analyze the concordance, false positive and false negative rates between circulating free DNA of the culture media compared to the results of the trophectoderm biopsy. MATERIALS AND METHODS: Observational, prospective and comparative study, conducted at Arcos Reproduction Center S.C. Nascere in november 2018. Couples with indication of preimplantation genetic diagnosis of aneuploidies undergoing In vitro Fertilization were included; carriers of structural anomalies or monogenic diseases were excluded and blastocysts with assisted hatching were eliminated. The response variables were the concordance, false positives and false negatives rates between trophoctoctoderm biopsies and culture media. Statistical analysis was performed with SPSS 25.0, using t-Student and chi-square tests with a value of p <0.05 significant. RESULTS: Informative results were obtained in 17 of the 20 culture media (85% successful global amplification); 70% on day 5 and 100% on day 6. The general concordance rate between trophectoderm biopsies and culture media was 68.7% (42.8% on day 5 and 88.8% on day 6). Regarding discrepancies, only 2 false negatives were observed in the culture media compared to the trophectoderm biopsy (14.2% on day 5 and 11.1% on day 6). There were 3 cases false positives (42.8% on day 5 and 0% on day 6). CONCLUSIONS: High rates of concordance were reached with the non-invasive genetic aneuploidy test, mainly in embryos on day 6.

Estrategia combinada en fertilización in vitro: mínima estimulación y transferencia de embrión único con diagnóstico genético. Experiencia de 3 años en dos centros de reproducción en México / A combined in-vitro fertilization strategy: minimal stimulation IVF, PGT-A and SET. Results from 3 years' experience at two IVF centers in Mexico

Resumen OBJETIVO: Evaluar los desenlaces de una estrategia combinada para fertilización in vitro: mínima estimulación ovárica, diagnóstico genético preimplantación para aneuploidias y transferencia de un solo embrión. MATERIALES Y MÉTODOS: Estudio de cohorte, retrospectivo, efectuado en dos centros de reproducción de México, en un periodo de tres años. Se incluyeron pacientes entre 25 y 45 años, en protocolo de fertilización in vitro, con mínima estimulación, diagnóstico genético preimplantación para aneuploidias (PGT-A) y transferencia de embrión único. El diagnóstico genético preimplantación se estableció mediante microarreglos y secuenciación de nueva generación (NGS). Para el análisis estadístico se integraron 5 grupos, según la edad de las pacientes: menores de 35 años; 35 a 37 años; 38 a 40 años; 41 a 42 años; y mayores de 42 años. Mediante estadística descriptiva se analizaron las variables numéricas y categóricas. RESULTADOS: Se analizaron 175 ciclos, en 125 pacientes (edad promedio: 39 años ± 5). Se obtuvieron, en promedio, 5 óvulos por ciclo. La tasa de fertilización fue de 86.5% y la de blastocisto por óvulo fertilizado de 50.7%. Se tomó biopsia para diagnóstico genético preimplantación para aneuploidias a 404 embriones. La tasa general de euploidia fue de 33%. Se efectuaron 69 transferencias de embrión único, con una tasa de embarazo por transferencia de 71%. La tasa de nacimiento por transferencia fue de 60.8% (42 nacimientos). CONCLUSIONES: La combinación de mínima estimulación, diagnóstico genético preimplantación para aneuploidias y transferencia de embrión único, es un procedimiento adecuado para alcanzar una tasa de nacimiento alta.

Abstract OBJECTIVE: To evaluate results of a combined approach in IVF, using minimal stimulation, preimplantation genetic testing for aneuploidy, and single blastocyst transfer. MATERIALS AND METHODS: Retrospective cohort study over a three years' period in two fertility centers in Mexico. A total of 125 patients were included, between 25 and 45 years old, with minimal stimulation IVF, preimplantation genetic testing for aneuploidy (PGT-A) and single euploid embryo transfer. PGT was performed using microarrays and next generation sequencing (NGS). RESULTS: A total of 175 cycles (mean age: 39 years old) were analyzed in 125 patients. On average, five eggs were collected per cycle; fertilization rate was 86.57%; blastocyst rate was 50.7% per fertilized egg. Only 33% of embryos were euploid. Pregnancy rate per transferred embryo was 71%. Live birth rate was 60.8% (42 births). CONCLUSIONS: A combination of minimal stimulation, PGT-A and single blastocyst embryo transfer can yield a high live birth rate.

Las contracciones en blastocistos humanos se correlacionan con aneuploidía, menor implantación y mayor tiempo de llegada a blastocisto: estudio retrospectivo con incubadora Time-Lapse

Objectives:To identify contraction (CT) patterns in human blastocysts using a Time-Lapse incubator and to correlate them with their ploidy status by PGT-A analysis, the time they took to reach blastocyst state, implantation rate and clinical pregnancy rate. Design: Retrospective cohort study. Interventions: From October 2016 through May 2018, 270 patients were evaluated, 912 embryos were cultivated in the Time-Lapse (Embryoscope™, Vitrolife) incubator; 778 of them were tested for aneuploidy using an NGS platform in a reference laboratory. Vitrification was performed according to the result of embryo development and awaiting the NGS result, followed by devitrification and transfer of a single embryo. Blastocyst contractions (CT) were determined using the EmbryoViewer (EmbryoViewer™ drawing tools) in order to obtain area, percentage of contraction and the various types of contractions, and these were compared with the NGS genetic study result. 182 transfers were performed to patients with an average age of 30.4 years, range 24 to 39 years. Finally, we obtained the correlation of implantation rate and clinical pregnancy of euploid embryos transferred in the program of assisted reproduction. Results: Embryos were separated into two groups according to contraction during their development: those that contracted (CT) and those that did not, named "only expanding" (OE). OE embryos were euploid in 58.3%, while 53.6% of CT embryos were aneuploid, with a statistically significant difference (p=0.029), meaning that OE embryos had higher chances of being euploid than CT embryos. Pregnancy rate was also higher in OE embryos (63.1% vs. 46.7% in CT embryos; p=0.012). CT embryos took longer to reach the blastocyst stage compared to OE embryos (p=0.004). Women's age had no correlation with embryo contraction. Conclusions: In this study, embryos who showed contractions had a higher chance of aneuploidy, a lower implantation rate, and required longer to reach the blastocyst stage. The simple fact of observing contractions in an embryo could be useful to decide to transfer another embryo without contractions. Further studies are necessary to prove these findings.

Objetivos. Estudiar los patrones de contracciones en blastocistos humanos mediante el uso de una incubadora time-lapse y correlacionarlos con su estado de ploidía por análisis PGT-A, el tiempo para alcanzar el estado blastocisto, la tasa de implantación y de embarazo clínico. Diseño. Estudio de cohortes retrospectivo. Intervenciones. Entre octubre 2016 y mayo 2018, se evaluó 270 pacientes; se hizo cultivo extendido de 5 a 6 días a 912 embriones en la incubadora time-lapse (Embryoscope, Vitrolife), y a 778 se les estudió para aneuploidía usando una plataforma NGS en un laboratorio de referencia. Hubo posterior vitrificación, según resultado del desarrollo embrionario y en espera del resultado del NGS, seguido de desvitrificación y transferencia de embrión único. Se determinó las contracciones del blastocisto (CTB) mediante la herramienta de dibujo del embrión EmbryoViewer (EmbryoViewer drawing tools), de manera de obtener el área, porcentaje de contracción y los diferentes tipos de contracciones, y se comparó los embriones con el resultado del estudio genético mediante NGS. Se transfirió 182 embriones en pacientes de 30,4 años promedio, rango entre 24 y 39 años. Finalmente, se correlacionó la tasa de implantación y embarazo clínico de los embriones euploides que fueron transferidos, en el programa de reproducción asistida. Resultados. Se separó los embriones en dos grupos de acuerdo a las contracciones durante su desarrollo, en aquellos que las tuvieron (CT) y aquellos que no, denominados 'solo expanding' (SE). Los embriones SE fueron euploides en 58,3%, mientras los embriones CT fueron aneuploides en 53,6%, con significancia estadística (p=0,012). Ello indica que la mayoría de los embriones euploides hacen 'solo expanding' durante su desarrollo, mientras que la mayoría de los embriones aneuploides (53,9%) hacen contracciones durante su desarrollo (p=0,029). Del mismo modo, la tasa de embarazo clínico de los embriones SE euploides fue 63,1% frente a 46,7% de los embriones CT, p=0.012. Finalmente, los embriones euploides CT tardaron más en convertirse en blastocistos tempranos que los embriones SE, p=0.004. La edad de la mujer no representó un factor para contracción embrionaria. Conclusiones. Los resultados obtenidos en este estudio muestran que los embriones que muestran contracciones, sin importar que tan intensas sean, están relacionados con mayor probabilidad de aneuploidías, menor tasa de implantación y ritmos de división lentos. El simple hecho de observar contracciones en un embrión podría ser útil para decidir transferir otro embrión que no las haya tenido. Se requiere más estudios para comprobar estos hallazgos.

Sorter de citometría de flujo: repercusiones en los indicadores clave de rendimiento de un laboratorio de reproducción asistida / Flow cytometry sorter: results in the key performance indicators of an assisted reproductive technology

Resumen OBJETIVO: Evaluar si la manipulación de gametos con sorter de citometría de flujo repercute negativamente en los indicadores clave de rendimiento de un laboratorio de reproducción asistida. MATERIALES Y MÉTODOS: Estudio descriptivo y retrospectivo, llevado a cabo en parejas a quienes se efectuó fecundación in vitro mediante inyección intracitoplasmática de espermatozoides (ICSI), con selección espermática, mediante un sorter de citometría de flujo, para selección de sexo. El estudio se efectuó en el New Hope Fertility Center de Guadalajara y Ciudad de México, de junio de 2014 a agosto de 2017. Los resultados se compararon con un grupo control seleccionado al azar. Se evaluaron los indicadores decisivos de rendimiento (KPI´s); tasa de fecundación normal, anormal (1PN, ≥ 3 PN) y fallida; tasa de degeneración posterior a ICSI; tasas de segmentación o división, blastocisto, implantación (segmentación y blastocisto) y recién nacido. Se utilizó la prueba t de Student para dos muestras y se consideró estadísticamente significativo el valor de p < 0.05. RESULTADOS: Se evaluaron 150 ciclos. Grupo 1: ICSI con selección espermática y sorter de citometría de flujo (n = 40); Grupo 2: ICSI sin sorter de citometría de flujo (n = 110). Los indicadores clave de rendimiento del grupo 1 disminuyeron; se reportaron tasas de fecundación fallida de 1.6%, blastocisto 17.4%, implantación en la segmentación 10%, implantación en blastocisto 14.2% y de recién nacido 14.5%. CONCLUSIONES: La manipulación de gametos con sorter de citometría de flujo reportó un efecto negativo en los indicadores clave de rendimiento del laboratorio de reproducción asistida, específicamente en las tasas de blastocisto, implantación de blastocisto y de recién nacido.

Abstract OBJECTIVE: To evaluate if the manipulation of gametes with a flow cytometry sorter has a negative effect on the key performance indicators (KPI´s). MATERIALS AND METHOD: Descriptive and retrospective analysis, in couples undergoing In a Vitro Fertilization (IVF) by ICSI, with sperm selection, using a flow cytometry sorter for sex selection. The study was conducted at the New Hope Fertility Center in Guadalajara and Mexico City, from June 2014 to August 2017. The results were compared with a randomly group without a flow cytometry sorter. KPI´s were evaluated; normal fertilization rate, abnormal (1PN, ≥3 PN), failed fertilization, ICSI damage rate, cleavage rate, blastocyst development rate, implantation rate (cleavage and blastocyst-stage) and live birth rate. A Student's t-test was made for two samples considering significant differences with p < 0.05. RESULTS: 150 cycles were evaluated. Group 1: ICSI with sperm selection by a flow cytometry sorter (n = 40); Group 2: ICSI without sperm selection (n = 110). Observing with statistical significance a decreased of the KPI´s of Group 1: failed fertilization rate (1.6%), blastocyst development rate (17.4%), implantation rate (cleavage-stage) (10%), implantation rate (blastocyst-stage) (14.2%) and live birth rate (14.5%). CONCLUSIONS: The manipulation of gametes with the flow cytometry sorter, has a negative effect on the assisted reproductive technology KPI´s; specifically, in the blastocyst rate, blastocyst implantation rate and live birth rate.

Effects of dynamic oxygen concentrations on the development of mouse pre- and peri-implantation embryos using a double-channel gas supply incubator system / 대한생식의학회지

OBJECTIVE: We aimed to evaluate the effects of different oxygen conditions (20% [high O₂], 5% [low O₂] and 5% decreased to 2% [dynamic O₂]) on mouse pre- and peri-implantation development using a novel double-channel gas supply (DCGS) incubator (CNC Biotech Inc.) to alter the oxygen concentration during in vitro culture.METHODS: The high-O₂ and low-O₂ groups were cultured from the one-cell to the blastocyst stage under 20% and 5% oxygen concentrations, respectively. In the dynamic-O₂ group, mouse embryos were cultured from the one-cell to the morula stage under 5% O₂ for 3 days, followed by culture under 2% O₂ to the blastocyst stage. To evaluate peri-implantation development, the blastocysts from the three groups were individually transferred to a fibronectin-coated dish and cultured to the outgrowth stage in droplets.RESULTS: The blastocyst formation rate was significantly higher in the low-O₂ and dynamic-O₂ groups than in the high-O₂ group. The total cell number was significantly higher in the dynamic-O₂ group than in the low-O₂ and high-O₂ groups. Additionally, the apoptotic index was significantly lower in the low-O₂ and dynamic-O₂ groups than in the high-O₂ group. The trophoblast outgrowth rate and spread area were significantly higher in the low-O₂ and dynamic-O₂ groups than in the high-O₂ group.CONCLUSION: Our results showed that a dynamic oxygen concentration (decreasing from 5% to 2%) had beneficial effects on mouse pre- and peri-implantation development. Optimized, dynamic changing of oxygen concentrations using the novel DCGS incubator could improve the developmental competence of in vitro cultured embryos in a human in vitro fertilization and embryo transfer program.

Influence of the Single Live Birth Outcome Following Transfer of Vitrified Blastocyst and Blastocyst Cultured from Thawing Cleavage Embryo / 中山大学学报(医学科学版)

[Objective]To compare the single live birth outcomes of blastocyst transfer between vitrified blastocyst and blastocyst cultured from thawing cleavage embryo,so as to choose the best scheme of blastocyst transfer.[Methods]Retrospective analysis of the single live birth clinical data of 1 037 vitrified blastocyst compared with 690 blastocyst cul-tured from thawing cleavage embryo undergoing frozen embryo transplantation(FET)from January 2014 to October 2016 was performed.Mail outcome were including gestational age,neonatal weight,proportion of male neonate,preterm birth rate,very preterm birth rate,low birthweight rate,very low birthweight rate,congenital anomalies rate.[Results]There were no differences between the two groups for gestational age,neonatal weight,proportion of Live birth,health baby and stillbirth(P>0.05). There were no differences in proportion of male neonate(AOR 1.07,95% CI 0.86~1.34),preterm birth rate(AOR 0.7,95% CI 0.49~1.01),very preterm birth rate(AOR 1.47,95% CI 0.55~3.96),low birthweight rate (AOR 1.38,95% CI 0.86~2.22),very low birthweight rate(AOR 0.76,95% CI 0.20~2.83),congenital anomalies rate (AOR 1.58,95% CI 0.66~3.76,P>0.05).[Conclusion]The blastocyst may be the preferable stage for vitrifying and transfer currently which can obtain good neonatal outcomes.