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Assisted reproduction techniques
Homologous and heterologous, 1st and 2nd level

Assisted reproduction is a set of various types of techniques, either medical or surgical, that can lead to conception and full term pregnancies in cases of sterility or infertility.

Unfortunately, in recent years, these have become increasingly common problems, and they are influenced not only by physical but also by psychological factors.
Our centre has long been involved in accompanying couples in the difficult search for an often long-awaited and deeply desired pregnancy.
The first step is to identify the particular causes that prevent conception and implement therapies aimed at curing them.
The diagnosis is carried by doctors at the clinic following specific examinations. In the event that is not possible to successfully treat the affected organ, the most suitable assisted reproduction tehniques are studied according to the specific problem.


One of the most common causes of female infertility is anovulation, i.e. the absence of ovulation, which may be accompanied by menstrual irregularities or even amenorrhoea.

These days, several drugs are available which can induce ovulation. They are becoming increasingly advanced and clinically efficient, which makes it possible to maximise therapy customisation in order to achieve the desired result.



IUI is the 1st level medically assisted reproduction treatment and involves moderate pharmacological stimulation of multiple ovulation. In certain circumstances it can be performed on a spontaneous cycle.
In order to follow the progress of folliculogenesis, ultrasound checks and on occasion hormone doses are required during treatment. If stimulation leads to the development of an excessive number of follicles (> 3), treatment can be interrupted to avoid the risk of multiple pregnancies.

In conjunction with ovulation, the male partner is asked for a semen sample: after appropriate treatment, the sperm is transferred into the uterine cavity via a catheter.
This technique is suitable in the event that infertility is due to the fact that the sperm are unable penetrate the cervical mucus and in the event of slight alterations in the seminal fluid.

Two weeks after insemination, a pregnancy test is carried out by checking for the presence in the blood of the β-HCG hormone produced by the embryo once it has been implanted. If positive, about three weeks later an ultrasound will follow to check for the presence of the gestational chamber.
Once identified, the pregnancy will be monitored over the following months.

In the event of a negative outcome, the couple will still consult with the doctor to assess how to continue.



It is a 2nd level treatment that consists of several stages:

Ovarian function stimulation

The first phase involves stimulation of the ovarian function in order to induce multiple follicle growth.
This is necessary because in vitro (i.e. in the laboratory) the probability of each individual oocyte being fertilised and developing into a foetus is lower than normal.
Multiple follicular growth allows more oocytes to be obtained per single ovulation, thus increasing the chances of achieving pregnancy.

Egg retrieval – PICK-UP

Follicles that are found to be sufficiently mature at the last stage of monitoring (diameter >16 mm) are aspirated and the follicular fluid obtained is immediately checked under the microscope for the presence of oocytes.
This is done using a simple needle aspiration technique while the patient is under sedation. The process is absolutely painless, transvaginal and aided by ultrasound.

In vitro insemination and embryo culture

Shortly after egg retrieval, the male partner is asked to produce a semen sample through masturbation.
After that, the sperm and oocytes will be placed in contact until the outcome of the insemination is ascertained.
Oocytes that show signs of normal fertilisation (oocyte with two pronuclei) are maintained in culture.
During this period the two pronuclei disappear, forming the zygote. This phase is followed by the first division of the cell, the moment in which we are in the presence of the actual embryo.
At our clinic, prolonged embryo culture is adopted up to the BLASTOCYST stage (5th - 6th day), a stage which can be reached in 90% of cases when there are at least 3 good quality embryos 48-72 hours after insemination, with results in terms of successful pregnancy of over 50%.

Embryo transfer – TRANSFER

On the day of the transfer, the embryos are transferred with a catheter into the patient's uterine cavity.
It is important to remember that the implantation process depends on both the quality of the embryos and the ability of the uterus to receive them.

Cryopreservation of remaining embryos

The remaining embryos that are not transferred and that show signs of viability are appropriately cryopreserved for possible subsequent cycles, without the need for re-stimulation.

Pregnancy tests

Approximately two weeks after the transfer, the ß-HCG dosage is analysed. This is a hormone produced by the embryo once it has implanted, and from this the outcome of the treatment can be ascertained.
If positive, about 20 days later a ultrasound will follow to test for the presence of the gestational chamber. Once observed, the pregancy will be monitored following months.
This technique is suitable in situations of tubal infertility, moderate to moderate-severe male infertility and in the event that intrauterine insemination fails.

Durata e passaggi trattamento Fivet



ICSI is a 2nd level IVF technique reserved for cases of severe seminal fluid alterations (very low sperm count and/or extremely reduced mobility), or after failure to fertilise oocytes through IVF.
The procedure is similar to IVF except that the oocyte fertilisation method involves the introduction of a single spermatozoid, via a micro-needle connected to a micromanipulator and under microscopic vision, directly into the oocyte.


The Assisted Medical Reproduction laboratory at the CMR clinic has been working with this particular type of embryo culture since 1998. It involves prolonged laboratory culture of the embryos until day 5 or 6, when they reach the blastocyst stage. They are then transferred into the uterus.
The blastocyst stage is reached in 90% of cases when there are at least 3 good quality embryos 48 to 72 hours after insemination, with results in terms of successful pregnancy of more than 50%.
Our laboratory has adopted the latest-generation GERI incubator with an INTEGRATED TIME-LAPSE facility, which, thanks to an integrated high-resolution video camera in each of the six incubation chambers, allows us to observe the development of embryos without disturbing them.

Each chamber is designed to contain the embryos of a single patient, eliminating the need to open the chamber containing other patients' embryos.
Geri enables PMA specialists to monitor every detail of embryo development conditions. It allows comprehensive observation of incubation conditions and specific safety mechanisms to ensure that an optimal culture environment is maintained at all times.
This state-of-the-art technology minimises stress on the embryos by ensuring optimal conditions for their growth and maximising pregnancy outcomes.

At CMR, the GERI incubator is used in 100% of the treatments performed.


Percutaneous Epididymal Sperm Aspiration

Testicular Sperm Aspiration

Both are sperm retrieval techniques in which spermatozoa are aspirated through a needle penetrating the skin, and then used in the event they are not detectable in semen.
PESA and TESA are necessary if the male partner presents azoospermia (lack of spermatozoa in ejaculate) due to an obstruction of the seminal tract.
In PESA, the sample is taken from the epididymis, while in TESA it is taken directly from the testicle.
With both techniques, egg retrieval can be carried out on the same day, with ICSI being performed with the sperm obtained.
It may be advisable, in some cases, to take a test sample before the stimulation cycle starts, in order to avoid unnecessary treatment.


Heterologous fertilisation is a medically assisted reproduction treatment available to couples with irreversible infertility or sterility problems.

This option became viable again in Italy in 2014, when the Constitutional Court, through sentence 162/2014, declared the ban on heterologous fertilisation imposed by Law 40/2004 to be unconstitutional.

This treatment involves the use of assisted fertilisation techniques, in which gametes (spermatozoa, oocytes or both) are provided by a donor from outside the couple.

We're talking about:
● heterologous fertilisation with egg donation
● heterologous fertilisation with sperm donation
● Heterologous fertilisation with egg and sperm donation (double heterologous)

Find out more by visiting the PGT-A Heterologous Fertilisation page



Nowadays it is very common to encounter couples resorting to medically assisted reproduction (MAR) programmes characterised by repeated failures.
This can manifest itself either in terms of failed pregnancies (implantation failures followingembryo transfer), pregnancies that have begun but ended in miscarriage, or pregnancies that are terminated due to the detection of a chromosomal pathology in the foetus, ascertained by prenatal diagnosis (amniocentesis or villocentesis).
In these patients, the reduced reproductive capacity is believed to depend on the presence of chromosomal aneuploidies in the embryos.
Aneuploidies are chromosomal alterations characterised by a greater or lesser number of chromosomes than the standard number, the incidence of which increases in proportion to a woman's age, resulting in a reduction in her reproductive potential.

The Preimplantation Genetic Test for Chromosomal Aneuploidies (PGT-A), previously referred to as PGT, makes it possible to assess embryo health in all couples undergoing in vitro fertilisationeven if they are not carriers of genetic diseases, allowing the transfer of embryos that are found to be free of chromosomal abnormalities on genetic analysis.
In these cases, the choice of embryos to be transferred into the uterus is based not only on their morphological appearance but also on their chromosomal arrangement, which reflects their potential to lead to a full-term pregnancy.
This test combines the use of IVF techniques with the latest genetics ressearch. Patients requesting access to pre-implantation diagnosis techniques will start medically assisted reproduction treatment, which involves the retrieval of oocytes to be fertilised with spermatozoa. Once fertilisation has taken place, embryos that have reached the blastocyst stage will be biopsied.
During the procedure, a number of cells will be extracted and their DNA will be analysed for chromosome alterations.

Embryos that are deemed normal upon genetic analysis, previously cryopreserved at the end of the in-vitro fertilisation and biopsy procedure, can be transferred into the uterus.
PGT-A improves the success rates of MAR treatments in women of so-called advanced maternal age, i.e. women aged between 36 and 43.
To find out more visit the website of the Genome Laboratory in Rome.



Preimplantation Genetic Testing of Monogenic Anomalies (PGT-M), previously referred to as PGD, is a technique that allows genetic diseases or chromosomal alterations to be identified in embryos at very early stages of development, generated in vitro by couples at high reproductive risk before their implantation in the uterus.

This test combines the use of IVF techniques with the most innovative research in the field of genetics. Patients requesting access to pre-implantation diagnosis techniques will begin medically assisted reproduction (MAR) treatment, which involves retrieving oocytes to be fertilised with sperm by means of ICSI. Once fertilisation has taken place, the embryos that have reached the blastocyst stage will be biopsied. During the procedure, cells will be extracted and their DNA will be analysed specifically in relation to the genetic disease to be tested for.

Embryos not affected by the genetic disorder, previously cryopreserved at the end of the in vitro fertilisation and biopsy procedure, can then be transferred into the uterus. After that it will be necessary to wait for the implantation outcome and to check for the presence of the gestational chamber. To date, diagnostic protocols exist for more than 200 monogenic, autosomal dominant, recessive or X-chromosome-related diseases. Genetic diseases that are very common in Italy, in which the PGT-M has a valid application, include Beta-Thalassaemia, Sickle Cell Anaemia, Haemophilia A and B, Dystrophy Duchenne-Becker Muscular Disease, Myotonic Dystrophy, Cystic Fibrosis, Spinal Muscular Atrophy (SMA) and X-Fragile Syndrome.

To find out more visit the website of the Genome Laboratory in Rome.



PGT-SR, on the other hand, allows the detection of structural chromosome abnormalities caused by the incorrect breaking or joining of chromosome segments, leadingultimately to disease.

There are many types of structural alterations:
translocations, deletions, duplications, insertions, ring chromosomes or inversions.


Vitrification of the patient's gametes and embryos is a procedure performed in the laboratory which allows their characteristics to be preserved over a period of time.

Vitrification involves the treatment of both gametes (where required) and embryos (for cases foreseen by law) with cryoprotectants and their subsequent immersion in liquid nitrogen at a temperature of -196° C. These very low temperatures keep the biological properties intact until thawing.

This procedure is far superior to the old freezing processes, as it prevents the formation of ice crystals that could damage cells.
It has been amply demonstrated by many studies that the survival rate of the vitrification process is about 97%.