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Intra Cytoplasmic Sperm Injection (ICSI)

Top Test Tube Baby Center in Hyderabad » Intra Cytoplasmic Sperm Injection (ICSI)

Gamete Handling Prior to ICSI

Fertilization by means of micromanipulation requires denudation of oocytes (i.e., removal of the surrounding cumulus and corona cells). This allows not only precise injection of the oocytes but also the assessment of their maturity, which is of critical importance for ICSI. Cumulus and corona cells are removed using the enzymatic procedure. Microscopic observations of the denuded oocytes include assessment of the zona pellucida and the oocyte, and the presence or absence of a Germinal Vesicle (GV) or a first polar body. ICSI is only carried out on metaphase II oocytes. Denuded and rinsed oocytes are incubated until the time of microinjection.

Routinely, sperm samples for ICSI are processed by density-gradient centrifugation [using silane-coated silica particle colloid solutions, enriching the number of motile and morphologically normal sperm cells needed for assisted reproduction.

Fertilization and Embryo Cleavage after ICSI

Post-fertilization, about 90% of 2-PN oocytes obtained by ICSI enter cleavage, resulting in multicellular embryos. Cleavage characteristics of the fertilized oocytes are evaluated daily. Normally developing, good-quality embryos reach the four-cell and eight-cell stage, respectively, on day 2 and in the morning of day 3 post microinjection. Numbers and sizes of blastomeres and the presence of anucleate cytoplasmic fragments are recorded. The cleaving embryos are scored according to equality of size of the blastomeres and proportion of anucleate fragments.

  • Type A (excellent quality) embryos do not contain a nuclear fragment.
  • Type B (good quality) embryos have a maximum of 20% of the volume of the embryo filled with anucleate fragments.
  • Type C (fair quality) embryos, anucleate fragments represent 21% to 50% of the volume of the embryo.
  • Type D (poor-quality) embryos have anucleate fragments present in more than 50% of the volume of the embryos.
  • These embryos cannot be used for the transfer to the patients. Embryos in the former three categories (type A, B, and C) are eligible for transfer.

Endometrial Preparation and Luteal Support

Progesterone supplementation is continued until the 12th week of pregnancy.