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Added: 09.08.2011

In vitro fertilization (IVF)

It is the most advanced method of assisted reproduction.  It is used in the following cases: obstructed tubes, endometriosis, abnormal sperm, or when the cause of infertility is unknown.  The treatment involves hormonal stimulation of the ovaries to receive several types of mature oocytes.  Shortly before ovulation (with the help of an ultrasound) the oocytes are collected, and after a brief final maturation in the incubator, properly prepared partner's semen is inserted.

After 18 hours a check up is needed to find out whether the oocytes were propertly fertilized, and the next day whether or not they began the process of partition.  If the partition progresses well, the embryos are transferred to the woman's uterus 3 days after the initial fertilization.  In some cases the culture of the embryos is extended  to 6-7 days or to the blastocyst stage.  Below, you can find a brief description of the various stages of the IVF:

 

Step 1-Hormonal stimulation of the patient (woman)

 

Woman goes through a hormonal treatment to stimulate the maturation of ovarian follicles and oocytes.  There are two types of stimulation: long protocol stimulation (a woman is given a hormonal product on the 21st day of the previous cycle), and short protocol stimulation (a woman is given a hormonal product on the 1st day of the current cycle).  The purpose of this hormonal product is to inhibit the secretion of sex hormones for a short period of time.  A proper stimulation begins on the third day of a cycle after a woman takes a medication that causes the growth of ovarian follicles.  The results of stimulation are controlled by an ultrasound.   A certain drug is administered 36 hours before the scheduled collection of cells to cause the final maturation of oocytes.

 

Step 2-Collection of oocytes

 

This procedure is performed in intravenous anesthesia under an ultrasound guidance.  By means of a special fine needle is inserted through the vagina, the doctor punctures the follicles and collects all the cells from both ovaries.  A light bleeding and abdominal pain may occur.  Those symptoms usually disappear after taking a painkiller.

 

Stage 3- Oocytes fertilization and culture of the embryos

 

Derived oocytes are placed in an incubator in specially prepared vessels with culture medium (nutrient fluid).  The oocytes are combined with sperm 3-4 hours after the maturation in the incubator.  After 18 hours it is usually known which cells were sucessfully fertilized.  2 days after the procedure the embryos reach the stage of 2-4 cells and on the third day-5-8 cells (blastomeres),  Typically at this stage the tansfer of the embryos is performed.  

 

Stage 4- Transfer of embryos into the uterus

 

In the final stage, usually on the third day after the collection procedure, the embryos are placed in the woman's uterus using a thin plastic catheter. Generally, 1-2 embryos are transferred at a time.  The transfer itself is completely painless.  From the day of the transfer, over the next 14 consecutive days it is necessary to take progesterone.  The progesteron positively affects the condition of the endometrium, and helps the implantation of the embryos in the uterus. 

 

Intracytoplasmic Sperm Injection (ICSI)

 

This method is a solution for men who have a small number of sperm in the semen, or for couples who have troubles with fertilization by classical IVF, despite good quality of eggs and sperm.  ICSI is performed under the guidance of a microscope.  A single sperm is inserted inside an egg with the help of a fine glass needle.  If a cell becomes fertilized, further culture and transfer is exactly the same as in the case of a classical in vitro fertilization.  

 

AZH (Assisted Zona Hatching)

 

Before conception and during an early embryonic development, the egg, and later the embryo are surrounded by a transparent shell (zona pellucia).  It plays an important role in the binding of sperm, and it also protects the embryo during an early development, and facilitates its transport through the oviduct.  After sperm enters the egg, the transparent shell hardens in its natural way; this prevents the polispermic fertilization to occur.  When the embryo reaches the blastocyst stage (5th or 6th day after fertilization), before implantation in the uterus, the shell becomes thinner, and then it breaks.  The embryo emerges from it, and it enters into a direct contact with the endometrium.  This process is called "hatching" of the embryo (embryo hatching).  If the embryo reamains in the shell the process of implantation does not occur.  The lack of hatching of the embryo may be caused by either a defective construction of the shell, and also its too strong hardening.  AZH is among the most used methods in the assisted reproductive techniques.  It involves a microsurgical  incision of the zona pellucida with an assumption of supporting the process of incubation.  The procedure itself is carried out using a simple and safe laser system.  In our clinic we use the Octax laser by MTG.  This is the infrared LED, which emits radiation with a frequency of 1.48 microns.  During the process of laser radiation, a gap (diameter of 5-10 um) is formed in the zona pellucida of the embryo.  Through the resulting gap, the embryo should escape from the zona pellucida and nest in the lining of the uterus.  This process is called an implantation.  This procedure is performed only on embryos selected for transfer.  The main advantage of laser technology, in comparision with other methods, such as an enzymatic method, is the lack of mechanical, chemical and thermal side effects acting on th embryo.  It is an efficient, fast and secure method.

Clinical observations have shown that after implementing AZH, where an incision was made in the zona pellucida, the percentage of implanted embryos was higher when compared with the results of transfering the embryos with an intact shell.  This procedure is particularly recommended to the following patients:

1. above 38 years of age-a tendency for too strong hardening of the zona pellucia increases with age

2. with elevated FSH

3.  with multiple failures in the treatment of infertility using assisted reproductive techniques

4. with recurrent loss of pregnancy (at least 2 identified during the interview)

5. with infertility conditioned by immune disorders 

6. with endometriosis

» View photo #1
» View photo #2

 

Embryo freezing (cryopreservation)

 

Embryos that are not used during the transfer are  frozen.  Frozen embryos can be stored for long periods of time with no detriment to their quality.   Frozen embryos can be used for future transfers.  In ths case, there is no need for a  subsequent hormonal stimulation of the patient.  If the first attempt succeeded and resulted in pregnancy, frozen embryos can be used to obtain subsequent pregnancies.  

 

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