- MEDICINE ATHENS MED4WOMAN ATHENS
- MEDICINE VOLOS MED4WOMAN VOLOS
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Obstetrician Gynaecologist
Doctor of Medicine, University of Athens Medical School
Panagiotis Polyzos, Gynaecologist Obstetrician, is active at the Institute of Life - IVF Unit of Iaso Maternity Hospital.
In Vitro Fertilization (IVF) is a series of medical procedures aimed at assisting fertility, facilitating conception, and preventing genetic abnormalities in the embryo. This procedure is also referred to as assisted reproductive technology.
IVF involves retrieving mature eggs from the ovaries, followed by fertilization with sperm in a laboratory setting. The resulting fertilized egg or eggs (embryos) are then implanted into the uterus. A full IVF cycle usually takes around three weeks to complete.
Today, approximately 10% of the reproductive population — which in Greece corresponds to around 300,000 couples — experiences infertility issues and seeks assistance from specialists through IVF treatment. IVF began its rapid development in 1978.
The main reason couples initially turned to IVF was due to damaged fallopian tubes, as the reconstructive surgeries available at the time had limited success. IVF bypasses the fallopian tubes entirely and transfers the pre-embryos directly into the uterus.
The IVF procedure is now performed using the most modern methods, achieving excellent success rates.
In a natural menstrual cycle, only one follicle typically matures and releases a single egg. However, to increase the chances of successful IVF, more than one egg is needed. For this reason, medications are administered to stimulate multiple follicles to mature, resulting in the production of a greater number of eggs.
Usually, the treatment that initiates follicular growth begins 14 days after the first injection, which is given on the first day of the menstrual cycle. The therapy consists of daily administration of gonadotropins, which stimulate the follicles to develop. The maturation of the follicles is monitored by the physician through ultrasound and hormonal testing (measurement of estradiol – E2 levels in the blood) on the fifth, seventh, ninth, and eleventh day of medication.
The dosage of the administered drugs is adjusted after each ultrasound and hormonal evaluation to achieve the best possible outcome. Through this approach, follicular growth is controlled, the timing of egg retrieval is determined, and ovarian hyperstimulation is prevented.
Pharmaceutical GnRH analogues resemble the natural hormone GnRH (Gonadotropin-Releasing Hormone), which is produced by the hypothalamus in the brain and stimulates the pituitary gland to secrete gonadotropins — the hormones responsible for controlling ovulation.
The natural GnRH remains active in the bloodstream only for a few minutes, whereas the analogues remain active for many hours. Initially, GnRH analogues stimulate the pituitary gland, but later they suppress its function in a paradoxical yet highly beneficial way. In IVF treatment, both of these mechanisms of the analogues are utilized.
Egg retrieval is performed 34–36 hours after the administration of hCG. The hCG injection supports the final maturation of the egg and its release from the follicular walls. The collection of the eggs is conducted using a needle that is guided via ultrasound through a transvaginal probe. The entire procedure typically takes 10–20 minutes, during which the patient is under analgesia or mild sedation.
The retrieved eggs are placed into special tubes and immediately transferred to the laboratory, where they are identified, counted, and placed in a specialized culture medium with temperature and chemical composition similar to the internal body environment. They are then placed in an incubator to continue maturing until fertilization takes place.
On the day of egg retrieval, the partner provides a semen sample in a sterile container. A period of abstinence of three to seven days is recommended beforehand. The sperm is evaluated for density and motility and is then prepared for fertilization.
The collected eggs are cleaned of surrounding cells and assessed for quality shortly before fertilization. Fertilization may occur using two main methods: conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI).In conventional fertilization, an appropriate concentration of sperm is combined with the eggs, and they are then placed in the incubator to allow fertilization to occur naturally.
In ICSI, a single sperm cell — selected for its optimal morphology and motility — is injected directly into the egg through a very fine needle. These eggs are then transferred to the incubator, where they remain until the next day, at which point embryologists check whether fertilization has taken place. The decision to use IVF or ICSI depends primarily on sperm quality. In cases where the sperm parameters are poor and cannot support successful fertilization through conventional IVF, ICSI is performed.
The day after egg retrieval (DAY 1), the eggs are examined under a microscope to determine whether fertilization has occurred. Successful fertilization is indicated by the presence of two pronuclei — one of maternal and one of paternal origin — inside the egg. The fertilized eggs are then returned to the incubator in a different culture medium, and the patients are informed accordingly.
The next step in embryo development is the division of the embryo into cells called blastomeres. The embryos are evaluated daily based on the number, size, and shape of the blastomeres, as well as the presence or absence of cell fragments.
On the second (DAY 2) and third day (DAY 3) after egg retrieval, embryos typically consist of around 4 and 8 blastomeres respectively. Embryo transfer usually takes place on day 2 or day 3. However, in some cases, embryo culture is extended until the fifth day after egg retrieval, when — after multiple divisions and further development — the embryos reach the blastocyst stage.
The embryo transfer procedure is very simple and painless, and it does not require analgesia or anesthesia. The number of embryos to be transferred depends primarily on their quality, and secondarily on whether a multiple pregnancy is desired. The maximum number of embryos allowed to be transferred is three in women up to 39 years of age and four in women aged 40 and above.
The embryos, along with a small amount of their culture medium, are drawn into a soft embryo transfer catheter, which is then passed through the cervix. Under ultrasound guidance, the embryos are placed into the endometrium near the top of the uterus (the uterine fundus).
The catheter is then removed and checked to confirm that all embryos have been successfully transferred. The woman typically remains lying down for about an hour and can then return home. After the embryo transfer, she continues progesterone therapy (Utrogestan or Crinone) daily until the pregnancy blood test. The pregnancy test is performed 14 days after egg retrieval.
If the result is positive, an ultrasound follows to confirm the presence of a gestational sac or sacs, and subsequently the detection of fetal heartbeats.
Cryopreservation is the process of freezing eggs, sperm, or embryos at sub-zero temperatures (-196°C) for future use. When eggs, sperm, or embryos are needed, they are thawed and used in the in vitro fertilization (IVF) process.
Embryo cryopreservation using the modern vitrification technique has increased the survival and quality rate of embryos after thawing to 95%.
The first child born from a cryopreserved IVF embryo is now 25 years old. The cryopreservation of human embryos has become part of daily clinical practice.
It was in 1983, in Melbourne, Australia, when the first child from a cryopreserved IVF embryo was born. The fertilized egg was frozen and, after thawing, transferred to the woman’s uterus, resulting in the birth of a healthy baby. Since then, more than 10,000 children have been born from cryopreserved embryos in Australia alone.
It is well known that embryo transfer has a certain probability of success, which depends primarily on the woman’s age. Therefore, if a single treatment cycle allows for multiple embryo transfers, the chances of achieving pregnancy increase significantly. Simply put, if more than one opportunity for embryo transfer arises from the same cycle, the likelihood of success rises dramatically. As the popular saying goes: “Persistence leads to victory.”
When a couple decides they are ready for the transfer of their cryopreserved embryos, the woman usually undergoes an artificial (controlled) cycle to prepare the uterus for embryo transfer. This artificial cycle involves oral medication, making the process very easy for the couple.
The maturation of the endometrium is monitored via ultrasound, and embryo transfer is carried out in the standard way.
The process of returning the embryos to body temperature takes approximately 2 hours and involves warming and rehydrating the embryos. During rehydration, the cryoprotectant agent is removed from the embryo cells by placing them in special salt-based solutions.
The reactivation of embryonic metabolism is evident as embryos continue to divide until they are transferred back into the woman’s body. Due to the stress of freezing and thawing, some cells may not survive, although the embryo can still remain viable and healthy.
It is also possible for an entire embryo to be lost. Pregnancy rates from frozen embryos are generally lower than those from fresh embryos. An embryo is considered to have survived thawing when at least 50% of its cells remain intact.
When 100% of the cells survive intact, the embryo has the same implantation potential as a fresh embryo. Overall, about 60–70% of embryos survive thawing, while 30–35% remain completely intact after cryopreservation.
Embryo cryopreservation is agreed upon with the couple for a period not exceeding 5 years. According to the law passed by the Hellenic Parliament on January 18, 2005, zygotes and fertilized eggs may be cryopreserved for up to 5 years, with the possibility of extending the period for an additional 5 years.
After the above period has expired, any zygotes or fertilized eggs that have not been used may either be allocated for research purposes or destroyed, subject to a decision by the relevant Authority. Cryopreservation of zygotes and fertilized eggs is carried out only upon written consent of the individuals depositing them, in accordance with Article 1456 of the Greek Civil Code.
Consent is permitted from the age of fifteen and above. Greek legislation also states that if sufficient embryos of good quality are available, the couple should not undergo a new IVF cycle before using their cryopreserved embryos.
To conclude on a positive note for couples undergoing IVF: current studies have shown that children born from cryopreserved embryos are absolutely healthy and equally comparable to children born naturally in the general population. Cryopreservation is a valuable tool in our effort to offer the much-desired child to couples facing infertility — a technique we should trust and utilize.
When the sperm produced during ejaculation is found in very low numbers (low sperm concentration — below 20 million/ml — and poor motility — below 50% progressively motile sperm), in vitro fertilization using the technique known as Intracytoplasmic Sperm Injection (ICSI) can provide a solution for these men.
With the ICSI technique, a single sperm cell is injected directly into the egg using a very fine needle. This approach effectively addresses low sperm concentration and poor motility.
In cases of azoospermia, where no sperm cells are found in the ejaculate, the situation becomes more complex. Azoospermia is found in approximately 10% of male infertility cases.
Until the mid-1990s, sperm donation was the only option for azoospermic men.
In cases of severe male infertility, ICSI is an IVF technique in which a single sperm cell is injected directly into the egg to achieve fertilization.
Today, sperm can be retrieved directly from the testicle and then used in the ICSI procedure to fertilize the partner’s eggs.
ICSI is employed because sperm retrieved through these methods are usually few in number and immotile or nearly immotile. Four different techniques exist for sperm retrieval, each with different names and acronyms. Every method has its own advantages and limitations, and not all are appropriate for every case. These techniques are the following:
MESA (Microsurgical Epididymal Sperm Aspiration)
Microsurgical aspiration of sperm from the epididymis. This method is indicated in men with obstruction of the ducts (tubules) that transport sperm from the testicles to the penis. The epididymis is accessed through a small one-centimeter incision in the scrotal skin. A surgical microscope is used to examine the very fine epididymal tubules.
A single tubule is opened, and its contents are collected and examined under a microscope to assess the presence and quality of sperm. The retrieved material is transferred to the laboratory for processing and, if an adequate amount is available, it is frozen for future use. It is important to cryopreserve any additional sperm, as a second IVF attempt is often required.
PESA (Percutaneous Epididymal Sperm Aspiration)
This is a non-microsurgical technique in which sperm is aspirated from the epididymis using a percutaneous needle. A needle is inserted directly into the epididymis with the hope that sperm will be found and collected. With this technique, only a small amount of sperm is typically retrieved, and it is rarely sufficient for freezing. PESA is not commonly recommended because it has lower success rates and a higher incidence of complications compared to other methods.
TESE (Testicular Sperm Extraction)
This is an open surgical procedure performed to collect testicular tissue, which can be done under local anesthesia and involves minimal risk of complications. A small piece of tissue is removed through a small incision in the scrotal skin.
The tissue is then placed into a special culture medium and divided into smaller fragments. Sperm cells are released from the seminiferous tubules, where they are produced, and subsequently separated from the surrounding testicular tissue. TESE typically yields an adequate amount of sperm of relatively good quality, which can be frozen and stored for future use. This technique is used in cases of both Obstructive Azoospermia (OA) and Non-Obstructive Azoospermia (NOA).
TESA (Testicular Sperm Aspiration)
Testicular sperm aspiration is performed using a fine biopsy needle that punctures the testicle to aspirate sperm. While this technique may yield sperm, the quantity obtained is usually low and often insufficient for freezing. The risk of complications, such as hematoma formation, is higher than with the TESE method.
In men with Non-Obstructive Azoospermia (NOA), sperm is rarely found in the epididymis; therefore, MESA and PESA are not recommended for these cases. TESE remains the most effective method of sperm retrieval for such patients. Testicular sperm retrieval techniques have now become part of routine clinical practice and have provided solutions for many couples facing infertility.
We now live in an era where the physician who previously could not find sperm in a semen analysis is no longer forced to tell the couple that they cannot have a child — instead, they can refer them to fertility clinics where effective solutions are available.
Fortunately, medical science has made tremendous progress in recent years, offering a range of alternative assisted reproduction options. Below, we analyze advanced techniques used in IVF. specialised IVF techniques.
Preimplantation Genetic Diagnosis (PGD) is a technique that can detect genetic abnormalities — either monogenic or chromosomal — in embryos before implantation, and therefore prior to pregnancy.
The first PGD case was reported in 1990 by Handyside and colleagues, who used the technique for embryonic sex determination. Only female embryos were selected for embryo transfer because the parents were carriers of a sex-linked genetic disease affecting only male offspring.
Since then, PGD has been applied to a wide range of genetic disorders (such as thalassemia, cystic fibrosis, muscular dystrophy, Down syndrome, Klinefelter syndrome, etc.), and the number of healthy children born following its use exceeds 1,000. This number is a testament to the accuracy and safety of the method, which is now offered in many IVF centers around the world.
PGD is directly linked to IVF centers because it is always performed in combination with an IVF cycle. Embryos must be formed in vitro (outside the body, in the laboratory) in order to be tested. Women undergo ovarian stimulation treatment, and at the appropriate time, egg retrieval is performed, followed by the following steps: fertilization, PGD testing on the fertilized embryos, and embryo transfer of the healthy embryos.
To determine whether an embryo carries a genetic disease or aneuploidy (a smaller or greater number of chromosomes than normal), one or two cells are removed — a procedure known as blastomere biopsy.
Biopsy is usually performed on the third day after egg retrieval and fertilization, when the embryo has reached the 8-cell stage. It may also be performed earlier (polar body biopsy) or later (blastocyst biopsy).
Biopsy is usually performed on the third day after egg retrieval and fertilization, when the embryo has reached the 8-cell stage. It may also be performed earlier (polar body biopsy) or later (blastocyst biopsy).
There are two types of diagnostic techniques, depending on the type of genetic abnormality being investigated. One is the PCR technique (Polymerase Chain Reaction), and the other is FISH (Fluorescence In Situ Hybridization).
The first detects genetic abnormalities in a specific gene (e.g., the β-thalassemia gene), while the second identifies abnormalities in the number or structure of chromosomes. Immediately after diagnosis, only the healthy embryos that have survived and continued to develop following biopsy are transferred into the uterus.
PGD is not applied only in infertile couples undergoing IVF due to fertility problems. It is also used in fertile couples who are able to conceive naturally but face a high risk of transmitting a genetic disorder to their embryos (e.g., thalassemia).
Other couples who may benefit from PGD include those with recurrent implantation failure or recurrent miscarriages. Additionally, the older a woman is when trying to conceive, the higher the likelihood of genetic abnormalities in the embryo. Therefore, PGD is also indicated for women of advanced reproductive age.
Overall, PGD can serve as an important tool in the hands of specialists to help select the “best and strongest” embryo.
By using PGD, only embryos that are genetically healthy and show the best developmental potential are selected, transferred, and ultimately implanted. In this way, the need for pregnancy termination at a later stage is avoided, anxiety regarding fetal health is reduced, and couples can focus positively on the pregnancy from the outset. PGD has already helped hundreds of couples at high risk of having embryos affected by genetic disorders bring healthy children into the world.
In combination with egg cryopreservation, another method of assisted reproduction is currently under research and development — the in vitro maturation (IVM) of immature gametes (oocytes and sperm). With this method, it is already possible — and will certainly be safer and more successful in the future — to mature gametes outside the body and later use them in IVF to create embryos and achieve pregnancy.
In women, this technique is particularly indicated in cases of polycystic ovary syndrome (PCOS), where hormonal stimulation to mature multiple eggs carries a high risk of ovarian hyperstimulation syndrome (OHSS). With IVM, ovarian tissue can be isolated and, without hormonal medication, the immature eggs collected can mature in the laboratory and subsequently be used for IVF once they reach the appropriate maturity level.
The method is also applied in women who, for medical reasons, must undergo oophorectomy (removal of the ovaries) or chemotherapy that may destroy ovarian function. These women can cryopreserve ovarian tissue and later use IVM to mature and utilize their own eggs when they wish to pursue pregnancy.
Additionally, women undergoing IVF with IVM may also mature any immature eggs collected during egg retrieval — thus increasing the number of embryos available and improving the chances of pregnancy.
For men, the in vitro maturation of sperm precursors may offer a solution to those who produce no sperm at ejaculation, and none are found in testicular tissue collected during surgical retrieval. These men usually have Non-Obstructive Azoospermia (NOA).
In such cases, testicular tissue can be cultured in the laboratory, allowing precursor germ cells found within it to differentiate into mature sperm cells. Through this technique, men who previously could only have children using donor sperm may, in the future, be able to father biological children with their own sperm.
One of the most important advances in in vitro fertilization is the freezing of eggs.
Today, the method is being applied, although it is still considered experimental. However, scientific progress will undoubtedly soon establish egg freezing as a routine procedure in IVF laboratories.
This method can help many women who, for various reasons, are not able to have a child at a given time.
One of the most significant reasons is medical: women who must undergo chemotherapy, which may destroy their ovaries and consequently their fertility, can freeze their eggs safely and, when they choose, thaw and fertilize them through IVF and transfer them to the uterus to achieve pregnancy.
Egg freezing can also provide solutions for women who, due to professional or social commitments or other personal circumstances, are at risk of exceeding their reproductive age without having had a child.
A significant advancement in science is the use of stem cells. Stem cells are undifferentiated cells that can potentially, through appropriate laboratory culture, develop into any type of cell (muscle, nerve, cardiac, liver, blood, etc.). These cells can be used in the treatment of various diseases such as leukemia, thalassemia, heart diseases, liver dysfunctions, etc.
Stem cells are the first cells of the embryo. In in vitro fertilization, on the fifth day after fertilization, the fertilized egg has divided and has formed a structure called the blastocyst. The blastocyst consists of cells at the periphery that will develop into the placenta and amniotic sac, and internal cells that will develop into the embryo. These internal cells are the stem cells.
Stem cells can be collected — although this requires the destruction of the blastocyst — and then cultured in order to produce a variety of cell types that can be used for the treatment of various diseases, usually affecting children of the same couple, since the likelihood of compatibility is higher.
The entire stem cell technology is currently undergoing rapid development and will certainly provide solutions to many problems in the future, not only diseases but also infertility, as stem cells may be used to produce gametes (oocytes or sperm). However, in the case of embryonic stem cells, the major ethical issue remains: the destruction of the embryo in order to obtain and culture the stem cells.
It is worth noting that stem cells are also found in adult individuals, as well as in the placenta and umbilical cord blood. Many parents already choose to cryopreserve umbilical cord blood and the placenta at the birth of their child for possible future use of their child’s stem cells.
The disadvantage of this type of stem cells, however, is that they are limited in number, difficult to isolate, and have a restricted — at least for now — pluripotency, meaning they do not differentiate as easily into various cell types as embryonic stem cells derived from blastocysts.
The IVF process is now performed using the most modern techniques and with particularly high success rates.
In the first stage, preparation takes place. For the woman, proper preparation is required, which includes hormonal testing, an ultrasound of the uterus and ovaries, and possibly an assessment of the cervix with a mock embryo transfer to ensure proper evaluation.
In many cases, there may already be indications of certain abnormalities, in which case a hysteroscopy should be performed to provide a complete clinical picture.
Apart from the woman who will undergo IVF, the man must also complete proper preparation before the procedure. Therefore, a semen analysis is required within 90 days before treatment. This allows for accurate evaluation of the sperm’s motility, morphology, and count.
The next step is the well-known process of ovarian stimulation. During this phase, multiple eggs are produced, in contrast with the natural monthly cycle where typically only one egg matures.
Throughout this phase, the woman undergoes hormone therapy under continuous monitoring by her doctor. This hormonal treatment may last from 14 to 28 days. During this period, the stimulation process is closely monitored so the doctor can evaluate progress and adjust medication dosages as necessary.
The next step is egg retrieval, which is explained in detail further below. Essentially, eggs are collected under light sedation, and the procedure lasts about 30 minutes.
Embryo transfer follows as the next step in the IVF process, during which the embryos obtained during treatment are placed back into the uterus. It is important to note that embryo transfer does not require anesthesia and can be performed immediately by the physician.
Finally, if everything has been completed correctly, blood tests are performed to measure the levels of hCG — the pregnancy hormone. The first pregnancy test is usually taken approximately 14 days after the last stage. However, this is always done in consultation with your gynecologist, who will provide the appropriate guidance.
It is a fact that this method requires a considerable amount of time and, for most couples, it represents something very important. Because the goal is clear and specific, in many cases the whole process involves stress, emotional pressure, financial cost, and much more. Most importantly, there is a general perception among the public that success rates are particularly high and that once the treatment is completed, everything will turn out exactly as expected.
For the psychological well-being of the couple, it is important to maintain realistic expectations and not to have hopes that exceed what a specialist has explained. Otherwise, if an attempt is unsuccessful, the psychological impact can be significant and may even lead to the couple deciding not to continue treatment.
In this effort, everyone involved is united. There is no doubt that the fertility specialist who supports you is on your side and shares the same goal. However, the most important aspect is that the doctor must first ensure the health of the woman and, overall, the health of the couple.
Therefore, Dr. Polyzos proceeds with a modern IVF approach with sincerity, compassion, respect, and a strong focus on the goal, achieving success rates significantly higher than the average. Based on the number of embryos transferred into the uterus — which is the final step of the entire process — the success rate reaches 40%–50%.
Throughout the IVF process, there are various factors that can influence the positivity of the outcome. In consultation with your gynecologist, these factors can be explained in detail, especially in relation to your specific situation.
However, some general factors that affect the success of IVF may include:
It is also important for the couple to understand that, beyond the above, achieving a successful IVF outcome requires strict adherence to the doctor’s instructions. Lifestyle and nutrition play a significant role, as do smoking and alcohol consumption, which can negatively affect treatment outcomes.
At this point, it is important to refer once again to the shared goal. The doctor and the couple become one team with a single objective: achieving pregnancy. A positive pregnancy test is a moment the couple will remember forever. The signs of a successful IVF can appear relatively soon.
Through blood tests, results can usually be detected as early as the first month. Classic pregnancy symptoms may occur, such as nausea, vomiting, constipation, and many others.
From the beginning of pregnancy, the uterus increases in size, preparing to host the embryo for the months ahead. Naturally, weight gain is expected and normal.
Finally, it should be noted that sexual intercourse does not affect the pregnancy as long as there are no complications that require restrictions.
We hope that all results are positive, so that a beautiful journey can begin with a joyful and growing family!
The issue of infertility is increasingly influenced by unprotected sexual activity, which has led to a significant rise in sexually transmitted infections such as chlamydia and gonorrhea. These conditions, if not diagnosed and treated early, can result in damage to the fallopian tubes — and consequently, infertility.
Additionally, the modern Western lifestyle, which often postpones pregnancy to later reproductive years — when peak fertility has already passed — makes IVF increasingly necessary.
Assisted reproduction techniques and methods are continuously advancing, and the future promises more solutions and higher success rates in IVF, even for couples who today must resort to options such as egg or sperm donation.
IVF is a complex treatment process, and its cost can vary significantly from one individual to another. Every couple has its own needs in terms of examinations and the techniques that are most suitable for their case.
To receive a detailed breakdown of IVF costs, a consultation with the doctor must first take place, during which a thorough evaluation will be performed and the cost will be determined.
At our clinic, we believe that IVF is a social good — a treatment to which all couples should have access. For this reason, we offer affordable pricing options. Contact us for more information!
In vitro fertilization (IVF) is a series of medical procedures designed to assist fertility, facilitate conception, and prevent genetic abnormalities in the embryo. This process is also known as artificial fertilization.
A complete IVF cycle typically lasts around three weeks. The duration may vary depending on the body’s response and the specific needs of each couple.
Throughout the IVF process, several factors may influence the success of the outcome. Your gynecologist can explain these in detail, tailored specifically to your case.
General factors affecting IVF success rates include:
Like any medical procedure, IVF carries certain risks. Despite some possible side effects, these risks are generally low and are effectively managed with modern techniques and close monitoring.
The age limit for IVF can vary depending on the country and clinic, but it is typically recommended for women under 42–43 years old, due to the natural decline in fertility with advancing age.
