A simple, less invasive stripper micropipetter-based technique for day 3 embryo biopsy
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METHODOLOGY ARTICLE
Open Access
A simple, less invasive stripper micropipetter-based technique for day 3 embryo biopsy Luciano Cedillo1, Azucena Ocampo-Bárcenas1, Israel Maldonado1, Francisco J. Valdez-Morales2, Felipe Camargo1 and Esther López-Bayghen1,3*
Abstract Background: Preimplantation genetic screening (PGS) is an important procedure for in vitro fertilization (IVF). A key step of PGS, blastomere removal, is abundant with many technical issues. The aim of this study was to compare a more simple procedure based on the Stipper Micropipetter, named S-biopsy, to the conventional aspiration method. Methods: On Day 3, 368 high-quality embryos (>7 cells on Day3 with 18 mm. Recombinant human Chorionic Gonadotropin (hCG) (Choragon 1000 IU, Laboratorio Ferring) was administered and oocyte retrieval was conducted 36 h after administration of hCG with ultrasound guidance. All 14–18 mm follicles were aspirated (typically 6 to 18), 6 to 14 oocytes were obtained with an average of 10.5 ± 2.5 oocytes per patient. Embryo culture
The partner's sperm of the female patient was prepared by density gradient centrifugation, and ICSI fertilization was performed as previously described [25]. The oocytes were all inseminated by ICSI at the same time after hCG administration (around 36 h), and fertilization was judged by the formation of two pronuclei, nineteen hours after insemination. Embryos were cultured in Global Total for Fertilization media (LifeGlobal) and incubated at 37 °C in 8% CO2, 5% O2, and 89% N2. Embryos were evaluated every 24 h and were assessed for development and quality. An Embryologist monitored and recorded all information about fertilization (rate = 67%), embryo development, embryo morphology,
Cedillo et al. Fertility Research and Practice (2016) 2:13
transfer and pregnancy for each cohort. Embryo morphology was assessed on Days 2 and 3 by considering the number of blastomeres, symmetry and granularity of blastomeres, type and percentage of fragmentation, presence of multinucleated blastomeres, and degree of compaction. After the biopsy, the embryo culture was extended to Day 5 (Global Total for Fertilization media, LifeGlobal) until Embryo transfer (ET), which was always performed with the optimal embryos. Blastomere removal
For both methods, only high quality embryos (66 h post insemination). Embryos with multinucleated blastomere may be biopsied if the other morphological parameters were determined to be ideal. All embryos were incubated until Day 5 to assess for embryo integrity and formation of the blastocyst after blastomere biopsy. For the Conventional Method, selected blastomeres were removed by the standardized protocol. Briefly, the embryos’ cell-to-cell adhesions were weakened by 5 min incubation in a Ca2+/Mg2+-free bicarbonate buffered GPGD medium (Vitrolife). Afterwards, the embryos were placed on a holding pipette (K-HPIP-1035, Cook Medical, Bloomington, IN). Using a Hamilton Thorne ZILOS-tk laser (1460 nm, 300 mW), an opening was created in the zona pellucida to expose a complete blastom
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