Expressed proteins and activated pathways in conditioned embryo culture media from IVF patients are diverse according to infertility factors.

OBJECTIVE: Given that the embryo culture medium secretome reflects the embryo development, we hypothesize that protein profiles are affected according to infertility factors, which can be responsible for detrimental embryonic developmental competence. The aim of this study was to screen the protein profile of conditioned embryo culture media in patients presenting deep infiltrating endometriosis (ENDO) and polycystic ovarian syndrome (PCOS) undergoing IVF, by proteomics approaches. The control group was constituted by tubal factor patients. METHODS: Patients underwent in vitro fertilization (IVF) treatment as routine and oocytes were fertilized by ICSI. The embryos were group cultured until day 3 of development, and after transfer the culture media were collected. For the proteomics analysis, two pools of samples were prepared for groups CONTROL and PCOS, and 4 pools of samples for group DIE. Samples were prepared to deplete high abundant proteins and followed evaluated by high throughput proteomics approach. RESULTS: The embryonic organ and tissue development were physiological functions activated, based on proteins identified in the 3 study groups of samples. The samples coming from DIE patients presented a high calcium activity and on the other hand, embryos coming from PCOS patients showed a decreased calcium action. Other pathways as grow factors through the EGF signaling pathway overexpressed in ENDO culture medium and protein kinase A in PCOS were also observed. CONCLUSIONS: Proteomic embryonic secretome will advance our knowledge of early embryogenesis and additionally could lead to improved selection of embryos for transfer warrants further investigation.

Does slow embryo development predict a high aneuploidy rate on trophectoderm biopsy?

The aneuploidy rates in expanded blastocysts biopsied on days 5 and 6 development were assessed in women undergoing IVF followed by array comparative genomic hybridization. This study included 1171 expanded blastocysts from 465 patients. Among the 465 patients, 215 and 141 underwent embryo biopsy on day 5 and day 6 (46.2% and 30.3%, respectively), and 109 underwent biopsy on both days 5 and 6 (23.4%). The cycles of 206 women were cancelled because only aneuploidy embryos were present (44.3%). The aneuploid embryos were classified according to the type as single, double or complex aneuploidy. No differences were observed in the distributions of these three categories according to the day of the biopsy. The aneuploidy rate was also evaluated according to maternal age, and was found to be higher in older patients; however, no differences in this rate were detected between embryos biopsied on days 5 and 6 according to maternal age. Biopsy was carried out when blastocysts reached the expanded stage. The embryos biopsied on day 6 had a higher rate of aneuploidy (69.9%) than those biopsied on day 5 (61.4%); however, the euploid embryos transferred had similar chances for successful and healthy gestation.

A Euploid Line of Human Embryonic Stem Cells Derived from a 43,XX,dup(9q),+12,-14,-15,-18,-21 Embryo.

Aneuploid embryos diagnosed by FISH-based preimplantation genetic screening (PGS) have been shown to yield euploid lines of human embryonic stem cells (hESCs) with a relatively high frequency. Given that the diagnostic procedure is usually based on the analysis of 1-2 blastomeres of 5 to 10-cell cleavage-stage embryos, mosaicism has been a likely explanation for the phenomena. However, FISH-based PGS can have a significant rate of misdiagnosis, and therefore some of those lines may have been derived from euploid embryos misdiagnosed as aneuploid. More recently, coupling of trophectoderm (TE) biopsy at the blastocyst stage and array-CGH lead to a more informative form of PGS. Here we describe the establishment of a new line of hESCs from an embryo with a 43,XX,dup(9q),+12,-14,-15,-18,-21 chromosomal content based on array-CGH of TE biopsy. We show that, despite the complex chromosomal abnormality, the corresponding hESC line BR-6 is euploid (46,XX). Single nucleotide polymorphism analysis showed that the embryo's missing chromosomes were not duplicated in BR-6, suggesting the existence of extensive mosaicism in the TE lineage.

Prospective randomized comparison of human oocyte cryopreservation with slow-rate freezing or vitrification.

OBJECTIVE: To compare cryopreservation of mature human oocytes with slow-rate freezing and vitrification and determine which is most efficient at establishing a pregnancy. DESIGN: Prospective randomized. SETTING: Academically affiliated, private fertility center. PATIENT(S): Consenting patients with concerns about embryo cryopreservation and more than nine mature oocytes at retrieval were randomized to slow-rate freezing or vitrification of supernumerary (more than nine) oocytes. INTERVENTION(S): Oocytes were frozen or vitrified, and upon request oocytes were thawed or warmed, respectively. MAIN OUTCOME MEASURE(S): Oocyte survival, fertilization, embryo development, and clinical pregnancy. RESULT(S): Patient use has resulted in 30 thaws and 48 warmings. Women's age at time of cryopreservation was similar. Oocyte survival was significantly higher following vitrification/warming (81%) compared with freezing/thawing (67%). Fertilization was more successful in oocytes vitrified/warmed compared with frozen/thawed. Fertilized oocytes from vitrification/warming had significantly better cleavage rates (84%) compared with freezing/thawing (71%) and resulted in embryos with significantly better morphology. Although similar numbers of embryos were transferred, embryos resulting from vitrified oocytes had significantly enhanced clinical (38%) pregnancy rates compared with embryos resulting from frozen oocyte (13%). Miscarriage and/or spontaneous abortion rates were similar. CONCLUSION(S): Our results suggest that vitrification/warming is currently the most efficient means of oocyte cryopreservation in relation to subsequent success in establishing pregnancy.

Extensive excision of deep infiltrative endometriosis before in vitro fertilization significantly improves pregnancy rates.

STUDY OBJECTIVE: We sought to compare the outcomes of in vitro fertilization (IVF) treatments in women with infertility-associated deep infiltrative endometriosis (DIE) who underwent extensive laparoscopic excision of endometriosis before IVF with those who underwent IVF only. DESIGN: Prospective cohort study. SETTING: Infertility clinic and private hospital in São Paulo, Brazil. PATIENTS: A total of 179 infertile patients younger than 38 years had symptoms and/or signs of endometriosis and sonographic images suggestive of DIE. INTERVENTIONS: After thorough counseling, 179 women were invited to participate in a prospective cohort study with 2 treatment options: IVF without undergoing laparoscopic surgery (group A, n = 105) and extensive laparoscopic excision of DIE before IVF (group B, n = 64). Ten women were lost to follow-up. The IVF outcomes were compared between the 2 groups. MEASUREMENTS AND MAIN RESULTS: In group B, patients had 5 +/- 2 (mean +/- SD) DIE lesions excised during laparoscopy. Patient characteristics in groups A and B, respectively, were: age (32 +/- 3 vs 32 +/- 3 years, p = .94), infertility duration (29 +/- 20 vs 27 +/- 17 months, p = .45), day-3 serum follicle-stimulating hormone levels (5.6 +/- 2.5 vs 5.9 +/- 2.5 IU/L, p = .50), and previous IVF attempts (1 +/- 1 vs 2 +/- 1, p = .01). The IVF outcomes differed between groups A and B, respectively, with regard to total dose of recombinant follicle-stimulating hormone required to accomplish ovulation induction (2380 +/- 911 vs 2542 +/- 1012 IU, p = .01), number of oocytes retrieved (10 +/- 5 vs 9 +/- 5, p = .04), and pregnancy rates (24% vs 41%, p = .004), but not number of embryos transferred (3 +/- 1 vs 3 +/- 1, p = 1). The odds ratio of achieving a pregnancy were 2.45 times greater in group B than in group A. CONCLUSION: Extensive laparoscopic excision of DIE significantly improved IVF pregnancy rates of women with infertility-associated DIE.

Punçäo testicular com agulha fina seguida de injeçäo intracitoplasmática de espermatozóide e gravidez: relato de caso / Fine needle biopsy of the testis followed by intracytoplasmic injection of the sperm and pregnancy: case report

Nos últimos anos, com o grande avanço das técnicas de reproduçao assistida, já se tornou possível a obtençao de gravidez utilizando-se apenas um espermatozóide, o qual pode ser injetado diretamente no interior do citoplasma do oócito (ICSI). Recentemente, a obtençao de espermatozóides através de punçao testicular com agulha fina nos mostrou que estes, apesar de imóveis e imaturos, sao capazes de fertilizar e resultar em gestaçao viável. Com isto, os pacientes portadores de fator masculino severo adquiriram uma nova esperança terapêutica. Apresentamos a seguir um caso de gravidez viável obtida com espermatozóide aspirado com agulha fina diretamente do testículo e ICSI.