IVF & ICSI at Genesis Fertility Center

Intracytoplasmic Sperm Injection (ICSI)

 IVF is a process in which the egg is fertilized by the sperm outside the body.
 In vitro means “in glass”.
 Embryos formed are transferred to the uterine cavity after a few days.

 ICSI is a specialized form of In Vitro Fertilization (IVF) that is used for the
treatment of severe cases of male-factor infertility.
 ICSI involves the injection of a single sperm directly into a mature egg.

History

 Attempts at in vitro fertilization date as back as 1953 however the first

pregnancy achieved through in vitro human fertilization of a human oocyte was
reported in The Lancet from the Monash University team of Carl Wood, John Leeton
and Alan Trounson in 1973, it lasted only a few days, biochemical pregnancy.
 There was also an ectopic pregnancy reported by Patrick Steptoe and Robert
Edwards in 1976.
 In 1977, Steptoe and Edwards successfully carried out a pioneering
conception which resulted in the birth of the world’s first baby to be conceived by
IVF, Louise Brown on 25 July 1978, in Oldham General Hospital, Greater
Manchester, UK.
 The first child born from gamete micromanipulation was a child in Singapore
born in April 1989.
 The technique was developed by Gianpiero Palermo at the Vrije Universiteit
Brussel, in the Center for Reproductive Medicine headed by Paul Devroey and
Andre Van Steirteghem.
 The procedure itself was first performed in 1987, It grew up to the pronuclear
stage. The first activated embryo by ICSI was produced in 1990, but the first
successful birth by ICSI took place on January 14, 1992.

Steps involved in IVF ICSI

Ovarian Stimulation
Ovaries are stimulated by hormones (FSH, HMG) to promote growth of follicles
containing eggs.
Monitoring of Follicular growth
The response of ovaries is monitored by ultrasound and/or blood tests. The dose of
hormones and days of stimulation are decided by the ovarian response.

Maturation and release of Egg

When the ovarian follicles have reached complete growth, induction of final oocyte
maturation is performed, generally by an injection of human chorionic gonadotropin
(hCG). hCG acts as an analog of the luteinizing hormone causes final follicular
maturation and release of oocyte and surrounding cumulus in the follicle. The
ovulation is imminent between 38 and 40 hours after a single HCG injection, The egg
retrieval is performed at a time usually between 34 and 36 hours after hCG injection,
prior to when the follicles would rupture.

Egg Retrieval (Ovum pick up)

This procedure is done under anaesthesia.

The eggs are retrieved from the patient using transvaginal sonography. The pickup
needle pierces vaginal wall under sonography guidance, reaches follicles. The
follicles are aspirated, and the follicular fluid is passed to an embryologist for
identification of oocyte.

Procedure of IVF

Semen is prepared for fertilization by removing inactive cells and seminal fluid by
centrifugation…. Semen wash.

Eggs and sperms are incubated in appropriate culture media.

Fertilization is checked after 16-18 hours.

Fertilized eggs are incubated for 48-72 hours

Procedure of ICSI

In the case of male factor involvement, intracytoplasmic sperm injection (ICSI) can be
done.

This involves following steps post oocyte retrieval

Egg Assessment

Cumulus cells surrounding egg are removed after treating them with Hylase enzyme

Eggs are graded as GV, MI and M II of which M II are mature eggs. M II oocytes are
considered for ICSI procedure.
Preparation of Sperm
After semen wash, motile sperms are selected for ICSI. Sperms are immobilized by
PVP or sperm slow (selection of mature sperms with fewer DNA strand breaks and
genetic abnormality) and selected for the procedure.

The procedure of Sperm Injection

It is done by specialized inverted microscope called Micromanipulator.

A holding pipette stabilizes oocyte by gentle suction created by a microinjector.

Hollow glass micropipette is used to cut the tail of immobilized sperm.

The oocyte is pierced and sperm is released in the cytoplasm.

The post-procedure oocyte is placed in culture and incubated, checked for signs of
fertilization after 16-18 hours.

Embryo Culture
Embryos are cultured in incubator till cleavage stage (Day 2-3) or Blastocyst stage

(Day 5-6).

The blastocyst has an advantage of higher implantation and ongoing pregnancy rate.

Embryo Selection

Embryo or oocytes are graded commonly by morphologic features, amount of cells,

evenness of growth and fragmentation before transfer.

Embryo Transfer

Selected embryos are placed in the uterine cavity by thin plastic catheter under
sonography guidance. A number of embryos transferred depend on availability and
National regulation.
Luteal phase support

Post-transfer hormonal support is given in the form of HCG and progesterone for 15
days after which a blood pregnancy test is done.
Vitrification

Good quality excess embryos are frozen and stored. These frozen embryos can be
thawed for subsequent cycles.