Effectiveness of Single Step Culture Media and Sequential Culture Media on Blastulation Rate
Effectiveness of Single Step Culture Media and Sequential Culture Media on Blastulation Rate
To compare the effectiveness of single step culture media and continuous uninterrupted
 culture media on blastulation rate
Sequential culture media are designed to mimic the changing environment in the ovary ducts
 and the uterus in vivo. However, there are some differences between in vivo and in vitro
 culture conditions. Hence, single step media are created which allow the embryo itself to
 choose the necessary nutrients while maintaining a more stable culture environment. Previous
 studies suggest higher blastulation rate in the single step media than the sequential media.

 This is an Randomized controlled trial (RCT) study comparing the blastulation rate of single
 step culture media and sequential culture media. Stimulation protocol is antagonist. Patient
 will be consulted on the day of ultrasound if they meet the inclusion criteria and sign the
 consent form. Then, the nurse will open the letter containing the type of media for that
 case. The media is prepared inside the lab a day before oocyte pick-up.

 After oocyte pick-up, the oocyte-cumulus complex (OCC) will be cultured for 2 hours at 37°C,
 6% Carbon dioxide (CO2) and 5% oxygen (O2). Meanwhile, sperm preparation will be performed
 using gradient technique. After 2 hours, OCC will be denuded and will be cultured for 1 hour
 until Intracytoplasmic Sperm Injection (ICSI). Oocytes after ICSI will be cultured in groups
 of 3-4 oocytes in the same conditions of 37 ° C, 6% CO2 and 5% O2. Fertilization check
 procedure is at 16-18 hours after ICSI. On day 3, the embryos will be evaluated in 66-68
 hours after ICSI, grade 1 embryos are identified with 7-9 blastomeres and fragmentation is
 from 0-10%. The patients will be informed about the embryo quality. If the embryos are not
 decided to have day 5 transferring, patient will be consulted to transfer on day 3.
 Blastulation rate will not be recorded.

 For the sequential culture media group, embryos will be transferred to Sequential Blast
 medium. For the single step culture media group, embryos will be transferred to the new
 culture dish with the same medium. On day 5, embryo quality assessment will be at 116±1 hours
 after ICSI. Blastocyst grading is based on the expansion, inner cell mass quality, and
 trophectoderm quality. Two good blastocyst will be transferred. After 11 days, beta-human
 chorionic gonadotropin (hCG) test will be required. If the beta-hCG result is positive,
 examination will be needed after 3 weeks and the luteal support will be given until 7 weeks.
Inclusion Criteria:

 -  Number of previous failure IVF cycles ≤2

 -  Using Gonadotropin-releasing hormone (GnRH) - Antagonist protocol for IVF

 -  Using GnRH for oocyte triggering.

 -  Read and sign consent form to participate in the study and have not been recruited for
 any other researches.

 Exclusion Criteria:

 -  Oocyte donation cycles

 -  Abnormal uterine.