Functional assessment of donated human embryos for the generation of pluripotent embryonic stem cell lines.

RESEARCH QUESTION: Can discarded embryos at blastocyst stage, donated to research because of genetic abnormalities and poor morphological quality, become a reliable source of human embryonic stem cell (HESC) lines? DESIGN: This study was consecutively conducted with 23 discarded embryos that were donated to research between February 2020 and April 2021. All embryos, except one, were morphologically evaluated and underwent trophectoderm biopsy for preimplantation genetic testing using next-generation sequencing (NGS), and then vitrified. After warming, the embryos were placed in appropriate culture conditions for the generation of HESCs, which was functionally assessed with immunofluorescence and flow cytometry for pluripotency capacity and spontaneous in-vitro differentiation. Cytogenetic assessment of the HESC was conducted with multiplex ligation-dependent probe amplification, and micro array comparative genomic hybridization. RESULTS: From the 23 embryos initially included, 17 survived warming, and 16 of them presented viability. Overall, the embryos presented poor morphological quality after warming. Only the previously untested embryo was capable of generating a new HESC line. Further characterization of this line revealed fully functional, euploid HESCs with preserved pluripotency, becoming a useful resource for research into human development and therapeutic investigation. CONCLUSIONS: None of the donated blastocysts with poor morphological quality in association with genetic abnormalities detected by NGS had the capacity for further in-vitro expansion to originate pluripotent HESC lines. This finding seems to provide extra support to genetic counselling on the suitability of this type of embryo for clinical use.

Early serum progesterone and prolactin analysis at day 9 of oocyte retrieval as a predictor of success in fresh ICSI cycles.

OBJECTIVE: To analyze progesterone and prolactin plasma levels nine days after oocyte retrieval and evaluate their correlation with pregnancy rates in in vitro fertilization cycles. To achieve pregnancy, several factors are analyzed before and during the in vitro fertilization cycle. Progesterone supplementation for adequate luteal phase support is indicated despite the presence of multiple corpus luteum in IVF stimulation cycles because of blockage caused by hypothalamic agonists and antagonists. The dosage of progesterone and prolactin on day 09 after follicular retrieval could function as a predictive marker of success in fertility treatments. METHODS: A retrospective study was performed using data from 238 patients submitted to intracytoplasmic sperm injection (ICSI) at a private infertility clinic from January 2013 to December 2015. Hormonal measurements were performed on day 09 after follicular uptake. The data was compared to assess the correlation between prolactin and progesterone dosages and pregnancy rates. RESULTS: The ICSI pregnancy rate was 40.8% (n=238). No statistically significant difference was observed when correlating the success of the procedure with the prolactin dosage (p=0.71). However, progesterone showed a significant difference (p=0.021). The cutoff point, indicated by the ROC curve fit according to which gestation would be identified, is 25.95ng/ml of progesterone. The sensitivity of this point is 61.9% and the specificity is 57.4%. CONCLUSION: Progesterone dosage may be one of the indicators of gestation on day 09 after follicular uptake. Such data can help physicians to monitoring and provides suitable early gestational care. More studies are needed to corroborate the data found.

A prospective randomized study comparing two commercially available types of human embryo culture media: G1-PLUS™/G2-PLUS™ sequential medium (Vitrolife) and the GL BLAST™ sole medium (Ingamed).

OBJECTIVE: To check the efficacy of two types of commercially available embryo culture medium: G1-PLUS™/G2-PLUS™ sequential (Vitrolife, Gothenburg, Sweden) and GV BLAST™ sole (Ingamed, Maringá, Brazil) with regards to fertilization, cleavage, blastocyst and pregnancy rates. METHODS: Prospective and randomized study conducted from March to July 2015, using the medical records of 60 patients submitted to Intracytoplasmic Sperm Injection techniques (ICSI). Data regarding the age of patients, together with fertilization, cleavage, blastocyst and pregnancy rates, were collected and compared in relation to the: G1-PLUS™/G2-PLUS™ sequential and GV BLAST™ sole mediums. The data were tabulated and compared using the Pearson's Chi-Square test (95% CI). RESULTS: There was no significant difference when comparing patients divided into higher and lower fertility age. No significant statistical difference was noted between the fertilization rates (P=0.59), cleavage (P=0.91), evolution to blastocyst (P=0.33) and total pregnancy (P=0.83) when comparing the embryos cultured in the different media analysed. CONCLUSION: We conclude that the G1-PLUS™/G2-PLUS™ sequential and GV BLAST™ sole mediums are equally effective with regards to fertilization, cleavage, blastocyst development and total pregnancy rates.

The First Ovarian Tissue Transplant between Monozygotic Twin Sisters Discordant for Ovarian function in Latin America.

Ovarian tissue transplant is an alternative to the cryopreservation of oocytes and embryos for the recovery of fertility and natural hormonal activity. The objective of this paper is to report on the first fresh ovarian tissue transplant between monozygotic twin sisters discordant for ovarian function, using the subcortical implant technique of ovarian tissue fragments, to take place in Latin America. A strip representing approximately a quarter of the cortical tissue was removed from the right ovary of the donor sister, cleaned, cut into small fragments and sent to adjacent room, where the receptor sister was concomitantly being prepared to receive the tissue graft. The ovarian fragments were placed under the cortical tissue onto a vascularized bed of the right ovary of the receptor sister. From 90 days postoperatively, the menstrual cycles of the receptor patient became regular with increased flow and longer periods, demonstrating normal hormonal activity and improved endometrial development. Attempts at spontaneous pregnancy, and the recovery of an oocyte followed by fertilization have not yet been successful. However, the ovarian tissue transplant between monozygotic sisters reported here clearly highlights the potential of the technique as a therapeutic option for the preservation of fertility.