G 1 plus g 2 plus

An intracytoplasmic sperm injection was performed for all patients with a standard micromanipulators (Narishige, Japan) and inverted microscope (Narishige, Nikon, Japan). Spermatozoa was microinjected with a × 200 magnitude in a drop of G-MOPS (Vitrolife, Sweden). Fertilized oocytes were placed in four-well dishes (Nunc, USA) in 500 μL G-1 PLUS (group Vitrolife). Zygotes and embryos were cultured in sequential medium (G-1 PLUS/G-2 PLUS, Vitrolife, Sweden). The culture was conducted in standard conditions (6% CO₂ and 5% O₂, 37 °C) using HeraCell 150 incubators (Thermoscientific, Germany).

Intracytoplasmic sperm injection (ICSI) was performed with micromanipulators (Eppendorf, Germany) and an inverted microscope (Leica DMI 3000B, Germany). Spermatozoa were microinjected under a 200 x magnitude in a drop of G-MOPS PLUS (Vitrolife, Sweden). Fertilized oocytes were placed separately in 20 µl droplets of G-1 PLUS Medium in Microdroplets Culture Dish (Vitrolife). Zygotes and embryos were cultured in sequential media (G-1 PLUS/G-2 PLUS, Vitrolife, Sweden). The culture was conducted in standard low oxygen conditions (6% CO₂ and 5% O₂, 37°C) using HeraCell 150 incubators (Thermoscientific, Germany). On day 3 the G-1 PLUS Medium was changed to the G-2 PLUS Medium. 15 µl of G-1 PLUS Medium from each droplet was stored at -70°C.