INFERTILITY TREATMENT

Laboratory Methods

The heart of every assisted reproduction clinic is the laboratory, as it plays a crucial role in the success of treatment. That is why at GYNCARE, we ensure not only that we have the most modern equipment available, but also that our embryologists and andrology laboratory technicians receive regular training and maintain high quality in their work.

Although laboratory methods are not covered by health insurance (with the exception of the ICSI contribution for UNION policyholders), their use can significantly increase the chance of successful treatment. You can consult our embryologists about the use of individual methods; they will automatically recommend the most suitable ones during an embryological consultation, taking into account your results and diagnosis.

Moderné embryologické laboratórium IVF centra s embryologičkou pozerajúcou do mikroskopu

Sperm Separation Methods

Mature sperm live only a few days. At the end of their life cycle, the incidence of DNA damage increases, accounting for about 15% of the sample. Even with a higher proportion of sperm with damaged DNA or other abnormalities, it is possible to separate such damaged sperm using one of the following separation methods:

Magnetic-activated separation allows for filtering out sperm with a high percentage of DNA fragmentation (damage) from a man’s ejaculate. Such sperm have an altered cell membrane capable of binding specific nanoparticles. These particles are then captured in a magnetic field along with the sperm they have bound to, while sperm without damaged DNA pass through the magnetic filter. Unlike IMSI and PICSI, sperm separated in this way can also be used for insemination (IUI).

This method mimics the natural mechanism of sperm selection. In this method, sperm must swim through special channels that filter out those with morphological abnormalities and changes in the cell membrane caused by DNA fragmentation. As a result, only sperm with optimal motility and good fertilization capabilities pass through this filtration method. This method significantly increases the success rate of pregnancy, reduces the risk of miscarriage, and is suitable for IVF, ICSI, and IUI. However, it cannot be used with thawed samples.

Polarization Microscopy – Oosight Spindle View

Oocyte maturity is determined by two important structures within the egg: the polar body and the meiotic spindle. An oocyte can be definitively considered mature if both of these structures are present. The polar body is a small sphere that clearly separates from the rest of the mature oocyte. Its presence can be checked even under a conventional microscope, and it forms earlier than the meiotic spindle. The meiotic spindle has such a delicate structure that it is not visible at all under a conventional microscope.

Polarization microscopy can reveal even fine details and structures of the oocyte, including the meiotic spindle. This means that embryologists can more accurately determine which oocytes are mature and which need further maturation in the incubator. This allows us to correctly time oocyte fertilization using the ICSI method, thereby increasing the chances of successful fertilization and development.

We recommend using this method for patients:

over 37 years of age
with low oocyte yield
who had inadequate embryo development in a previous IVF cycle despite normal spermogram parameters

We also recommend it in cases of:

High yield of immature oocytes
oocytes that do not have an optimal microscopic appearance
Use of thawed oocytes
In cases of a low fertilization rate in a previous IVF-ICSI cycle

ICSI (Intracytoplasmic Sperm Injection)

The principle of this micromanipulation method involves fertilizing a mature oocyte by injecting a sperm directly into the oocyte using a fine needle. The sperm is selected based on its morphology and motility, or one of the PICSI, IMSI, MSS, or MACS methods can be used prior to ICSI. This method can only be applied in in vitro fertilization (IVF) and eliminates male infertility factors, especially low sperm motility and concentration (count). ICSI significantly increases the chances of fertilizing the highest possible number of oocytes, with fertilization success rates reaching around 80%.

We especially recommend this method for couples in cases of:

Spermogram abnormalities (poor results)
Failure of previous IVF treatment
If a lower number of oocytes was retrieved during collection
Use of frozen sperm
Use of donor oocytes
If genetic testing of embryos is planned

This method utilizes the fact that only mature sperm have binding sites for hyaluronic acid on their surface, which is part of the oocyte’s outer layer. In this method, sperm are placed in a hyaluronan-containing environment. Mature and healthy sperm have receptors on their surface that allow them to bind to this hyaluronan, enabling their selection for subsequent oocyte fertilization using the ICSI method. The method is recommended for patients:

with pathological spermogram parameters (however, it is technically not feasible with very low sperm motility in the ejaculate)
with impaired embryo development in previous IVF cycles
in cases of advanced paternal age

Intracytoplasmic injection of morphologically selected sperm is based on maximum microscopic magnification of the sperm, allowing the embryologist to evaluate even minimal defects in sperm morphology. This method cannot be used with a very low sperm count in the ejaculate.

This is a special medium (solution) designed to distinguish live sperm from non-live ones. It is most commonly used for sperm obtained through surgical retrieval – MESA/TESE, as well as in cases of necrotic sperm findings. Its composition temporarily accelerates otherwise immotile or very slowly moving sperm. Their activation allows the embryologist to avoid overlooking healthy sperm that might appear dead at first glance due to their immobility. These live and viable sperm, which have a chance of retaining their ability to fertilize an oocyte, are then used for fertilization via the ICSI method.

When an oocyte meets a sperm, oocyte activation is triggered, initiating the process of fertilization and embryo development. This activation is caused by sperm factors that lead to increased calcium production within the oocyte. If the sperm lacks these factors, or if the oocyte does not react correctly to them, the activation process may not occur, and fertilization may fail. Assisted activation uses a calcium ionophore, into the solution of which the fertilized oocyte is placed for approximately 15 minutes. It is then cleaned and placed in an incubator for culture. Assisted oocyte activation is particularly suitable if ICSI fertilization failed in a previous cycle or when using sperm obtained from testicular tissue via microsurgical retrieval (MESA/TESE).

Embryo Culture

In natural conception, approximately 5 to 6 days pass from oocyte fertilization to embryo implantation in the uterus. During this time, through cell division, the embryo reaches the blastocyst stage, and only after reaching this stage does it leave the oocyte’s outer layer to implant. However, the development of some embryos stops during this period for various reasons – most often genetic. In such a case, the embryo does not implant, leading to a spontaneous miscarriage that is indistinguishable from a regular menstrual period.

In assisted reproduction via IVF, the oocyte is fertilized in the embryology laboratory. If fertilization is successful, embryologists monitor the embryo’s cell division for the next three days. On the third day, the embryo is transferred to the uterus (embryo transfer). However, at this point, it is not yet certain whether embryo development will stop or if the embryo will reach the blastocyst stage, with cell division most commonly ceasing between 3. and the 4th day of development. Unfortunately, health insurance companies only cover 3-day culture within an IVF cycle. However, patients can choose the option of extended culture.

Embryo culture takes place in a special incubator that maintains an optimal atmosphere for proper embryo division. This incubator has multiple chambers, each divided into several sections. Embryos from a single patient can be stored in each section. Whenever an embryologist checks the development of embryos for a patient, they must open the incubator chamber. With each such opening, the optimal atmosphere is briefly disturbed for the embryos in all four sections of the chamber.

With individual embryo culture, each patient has an incubator chamber exclusively for her own embryos. This ensures a stable atmosphere in the incubator, which improves early embryonic development and thus enhances the chances of embryos implanting in the uterus and further developing.

With extended culture, embryos can be monitored up to the fifth or even sixth day. This allows us to transfer only those embryos that have reached the blastocyst stage, thereby significantly increasing the chances of a successful treatment cycle leading to pregnancy. Extended culture is standardly used and highly recommended also for freezing (vitrification) of remaining embryos, as only embryos that have reached the blastocyst stage are frozen. These also tolerate freezing and subsequent thawing better.

In extended culture, embryo development is monitored manually – by removing them from the incubator and placing them under a microscope. This is done every day of culture. Thanks to the use of the EmbryoScope+ time-lapse incubator, we can continuously and much more thoroughly monitor embryo development throughout the entire culture period. This is made possible by a built-in camera that captures each embryo individually at regular intervals of several minutes. The embryos are cultured and monitored individually in their own chamber. As a result, they do not have to leave the incubator environment, which helps maintain optimal conditions for their development. The result is a time-lapse video, which allows the embryologist to analyze the course of development and successfully detect any anomalies during division. This makes it possible to select the most suitable embryos with the highest implantation potential for embryo transfer or freezing, thereby helping to increase the probability of a successful cycle.

The use of time-lapse monitoring is especially recommended for couples:

who have experienced repeated implantation failure
who are interested in genetic testing of embryos
where the woman is over 35 years old
where a severe andrological factor exists
in cases of higher oocyte and embryo yield, it allows for better selection of the most suitable ones for transfer

Assisted Zona Hatching and Laser-Assisted Zona Pellucida Thinning

Up to a certain size, the embryo develops protected by a firm outer layer (zona pellucida). This layer ruptures on the 5th-7th day after fertilization due to embryo growth, allowing the embryo’s cells to attach to the uterine lining – known as implantation. In some cases, the outer layer is too rigid, preventing its rupture, and thus the embryo cannot exit and implant, which prevents pregnancy. Such a layer is usually somewhat thicker, allowing embryologists to indicate the need for assisted hatching or thinning, but these procedures can also be performed preventively.

The essence of this procedure is to disrupt the outer layer using a special laser and create a small opening just before embryo transfer to the uterus.

In this case, the zona pellucida is only thinned using a laser beam to facilitate the embryo’s exit.

Both of these procedures are safe and without risk to further embryo development. Considering preventive AZH or LAZT is especially appropriate:

for older women who tend to have oocytes with a thicker outer layer
in cases of repeated implantation failure
when using frozen embryos

The LAZT method is also suitable for lower quality embryos or embryos without extended culture.

EmbryoGlue

The transfer of an embryo to the uterus (embryo transfer) and its subsequent successful implantation in the uterine lining are among the most crucial steps in infertility treatment using IVF methods. EmbryoGlue is a culture medium (solution) specifically developed for embryo transfer, containing a high concentration of certain substances (hyaluronan and recombinant human albumin) that help mimic uterine conditions and increase embryo adherence to its lining. This demonstrably increases the chances of a successful pregnancy, as shown by an international study, according to which the clinical pregnancy rate increased from 41% with the use of a standard culture medium to 49% with the use of EmbryoGlue (Bontekoe et al. Adherence compounds in embryo transfer media for assisted reproductive technologies (Review). The Cochrane Collaboration 2014).

Price list

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Treatment Options

All these methods can be used in infertility treatment via IVF and in treatment using donor oocytes. Sperm separation methods can also be used in intrauterine insemination, and EmbryoGlue in donor embryo treatment.

IVF