More additional IVF lab techniques, the better IVF success rates?

Yadira Pallas Galvez, MSc
Senior Clinical Embryologist , UR Vistahermosa

Failed IVF Cycles, Genetics PGS / PGT-A, Male Factor, Success Rates

More additional IVF lab techniques, the better IVF success rates? The medical evidence
From this video you will find out:

In the webinar, the participants had a chance to find out whether extra IVF lab procedures like

  • ICSI
  • IMSI
  • PICSI®
  • hbIMSI
  • Assisted Hatching
  • Embryo Glue
  • PGT-A (formerly called PGS)
  • PGT-M and PGT-SR (formerly called PGD) and other
  • influence the outcome of IVF treatment and boost your success rates.

IVF Laboratory procedures vs better IVF success rates

IVF and IVF with donor eggs – the more additional procedures you have, the better success rates. Is this true? What is the expert’s opinion and medical evidence? Watch the recording from live webinar and Q&A session with Dr. Yadira Pallás Gálvez, an experienced embryologist from the Reproduction Unit of Hospital Clínica Vistahermosa (PreGen) in Alicante, Spain.

Do additional lab techniques mean higher IVF success rates?

There is much conflicting and controversial information surrounding additional IVF procedures leaving some patients to approach these laboratory “add-ons” with caution. Questions are inevitably raised regarding the actual techniques used and whether an increase in additional treatments will actually provide an increase in pregnancy rates.

In this webinar, Dr Yadira Pallas Galvez, senior clinic embryologist at PreGen in Alicante, Spain, describes the basic IVF procedure, outlining the various additional techniques currently used and explaining when further treatment options should be recommended for patients.

The object of IVF is to fertilise eggs and create embryos outside of the body. To optimise the number of potential embryos, the first step of IVF is to increase the amount of natural eggs produced by the body, this is known as ovarian stimulation. During this stage of the IVF process, the follicles (the cavity which surrounds the oocyte) are regularly checked by ultrasound to establish the optimal time for their retrieval. An egg collection is then carried out, under sedation, and once any oocytes have been obtained, they are placed in a special incubator until the sperm has been properly prepared in the laboratory.

Finally, either IVF or ICSI is performed using the fully mature eggs. For IVF the sperm and egg are placed together in the incubator, whereas with ICSI one specially selected spermatozoa is injected into one egg; this micro injection technique is usually used in cases of male factor infertility.

After 18-20 hours the first phase of fertilisation take place and embryo cultivation begins. As they grow, all embryos are classified by the embryology team according to their development. This grading helps embryologists determine when the transfer will take place and which embryo/s should be transferred or vitrified (frozen).

Embryo transfers are not painful and do not require sedation. The embryos are inserted into the uterine cavity using a catheter and ultrasound guidance. The number of embryos transferred depends on the patient’s history, the embryonic quality and medical advice; clinics aim to transfer a single, good quality blastocyst (five-day embryo) to achieve the best results however, this isn’t always possible.

So, what else can patients do to try to achieve a good quality blastocyst for transfer?

Where male infertility is a factor, Dr Galvez would recommend a MACS test. All spermatozoa have a limited viability and through the process of Magnetic Activation Cell Sorting (MACS), any apoptotic (dying cells) sperm become separated. This separation of the gametes (sperm) enables embryologists to only use sperm which is alive and healthy for the fertilisation process. Apoptotic sperm are likely to include a higher amount of DNA fragmentation which can result in abnormal embryo development. DNA fragmentation cannot be detected when using basic microscopic methods. Dr Galvez suggests MACS testing for patients who are suffering from recurrent implantation failure, a low fertilisation rate or poor embryo quality, and when male age is increased.

Research into genetics has opened up huge opportunities in the field of assisted reproduction and with the introduction of preconception genetics testing, medical teams now have the possibility of increasing the chances of a genetically healthy pregnancy. Specialists would typically test for any recessive diseases, these include conditions such as Cystic Fibrosis, Spinal Muscular Atrophy and Beta Thalassemia, which occur when both parents are determined to be carriers of mutations for the same disease. Genetics testing allows medical teams to review the compatibility of couples to determine whether they are genetically suited. For couples who are incompatible further tests and options are available and the use of donor gametes may be suggested as an alternative approach.

Genetic screening can also be carried out on embryos by using PGT-A (pre-implantation genetic testing for aneuploidies). Aneuploidy is the presence of an abnormal number of chromosomes in a cell and is the most frequent cause of implantation failure, miscarriage and congenital anomalies (birth defects). The risk is known to increase with maternal age. Pre-implantation screening can improve implantation and pregnancy rates and maximises the opportunity of transferring only euploid (chromosomally equal) embryos. The aneuploidy testing follows an embryo biopsy, where a small number of cells and removed and then analysed.

The use of time lapse technology is another additional treatment regularly offered to patients, and one which PreGen clinic has recorded a 20% pregnancy increase following their usage.

Time lapse is an optical system within the incubator which allows embryologists greater opportunity to study embryonic development throughout the entire cultivation period.

When time lapse imaging is not used by clinics, embryologists have to physically remove the embryos at least every 24 hours so that their development can be analysed. Removing the embryos from their incubator exposes them to microscopic lights, temperature and humidity changes and human manipulation. Time lapse incubators limit this exposure and instead offer constant, optimal cultivation conditions. As embryologists are afforded around the clock embryo observation, time lapse technology helps them to select only those with the highest implantation potential for transfer and freezing.

Assisted hatching and embryo glue are two other laboratory add-ons which can be performed to aid implantation. In assisted hatching a minor defect is created in the embryo membrane which potentially increases implantation potential. It would typically be recommended for embryos that have morphological issues that might make implantation more difficult. Embryo glue is used as a transfer medium and is enriched with everything an embryo needs to help it implant. As of yet, no clear data has been collated to confirm whether these two techniques have a significant impact on implantation.

Dr Galvez advises that it is important for patients to remember that these are additional treatments and are not always necessary, or applicable, for everyone. Clinics must assess all patients individually and only suggest extra lab techniques which will make a difference in each unique set of circumstances.

- Questions and Answers

So, are there any indications to use EmbryoGlue?

We think that the use of embryo glue should be general and used all the time, as it always helps. There is not anything in the embryo glue that could damage the cycles.

Are there any publications about embryo glue proving that it’s better to use it?

Yes, that’s why we use it. There is medical evidence and bibliographies reporting the embryo glue use.

Do you have experience with PRP?

We do not use it at the moment, as there is no medical evidence for it.

Any specific add-on that you would not recommend?

All of them are useful, only when medically necessary. It doesn’t work in a way that “the more you do, the better”—there has to be a medical indication for it.

We have one blastocyst (day 5) that is just being frozen. Our clinic have recommended maybe doing a mild stimulation/natural cycle in the hope to transfer a fresh embryo on day 3 and transfer the frozen on day 5. Could this increase our chances?

We normally do not use it, so we do not recommend it. We think that it is better to transfer all the embryos on the same day—there is no evidence to suggest the chances will be greater to transfer fresh embryos on one day and then frozen embryos later, so we cannot answer this question. In our opinion, you can transfer two of them on day 5.

My husband has high sperm fragmentation and we are using donor eggs and his sperm. We have achieved one euploid embryo which has been looked at for chromosome abnormalities. Could his sperm fragmentation still cause issues with implantation or live birth?

Seeing as the embryo has already been screened, and the sperm fragmentation has been solved (?-27:56), if the embryo is euploid, it does mean that there is sperm fragmentation and it’s okay, so implantation and live birth should be good.

I’ve had the scratch three times but no implantation so far, do you recommend the scratch?

We normally do the scratch when there is a medical indication for it. The scratch helps implantation, so it depends on your doctor—if they think that you need it, then, of course, you should have it.

Have you had a patient who after many IVF failures with own eggs had success after using an additional procedure?

This can happen with PGS, when you screen the embryos genetically because probably in the previous IVFs we don’t know if the embryos were genetically normal or not, so after performing the PGS, we are sure that the embryos which we are transferring are viable to have implantation and then a baby.

Would you recommend to use an AH on fresh cycle- fresh embryos as I know there are clinics doing AH for every fresh embryo before the transfer.

We do not recommend to do it generally. There are some embryos which have implantation capacity, so you don’t have to do assisted hatching, and there are others where assisted hatching does need to be done, so these are the cases where we perform this technique.

What do you think about testing the lining of the womb to look at endometrium receptivity? Can this improve the chance of achieving pregnancy?

Yes, it definitely helps. If the womb is not ready, or if it is not prepared or receptive, it’s very difficult to achieve a pregnancy. So, all techniques to measure the womb or to measure the endometrium receptivity are quite important to be successful.

Do you perform hormone blood tests prior to transferring the embryo from egg donation?

We only deal with the endometrium receptivity because it’s very difficult to measure the hormones because the hormones that we are giving are artificial.

Embryo monitoring—do you recommend it for every patient? Is it really something that can help or is it just a marketing strategy so we can have the video of our embryos while developing?

We do not consider it a marketing issue—we think time-lapse embryo monitoring is one of the most important techniques because it gives the clinics and laboratories a lot of information about what is happening inside the incubators. In a normal incubator, you should remove the embryos every day to check how they are, and you can miss lots of things, but when you do embryo monitoring, you have everything recorded, and you have much more information, which can help the embryologist to decide which is the best embryo to be transferred. We consider it to be quite important in a clinic to have an embryo monitoring system, and all patients can benefit from it, as you don’t need a medical indication for it to be used.

Can you avoid using ICSI process with egg donation? I have heard it can cause future problems.

There is no medical evidence for that. We choose whether or not to do it depending on the quality of the sperm, not on the source of the eggs. It helps with fertilisation when necessary, and we don’t have any medical parameters to confirm that it could cause any problems—we think it can help to achieve a pregnancy. The sperm sample will determine if we need ICSI.

Do you think testing for immune issues and NK cells is important if a lot of implantation failures have happened?

Yes, of course, it is important because when you have had implantation failures or miscarriages, you should have a battery of tests—you should have a protocol of implantation failure cases. These tests include immune tests, NK cells and also others, so yes, it can give us the origin of the problem.

What do you think of PICSI? Can you explain it and its benefits for high sperm fragmentation issue?

When we have high sperm fragmentation, what we do is MACS (magnetic-activated cell sorting)— it has a lot of medical evidence to support it. It removes most of the apoptotic sperm, that means the sperm that is damaged. After that, we do the PICSI, when we have a camera with up to 400% magnification. With that, we have very good results using the MACS + PICSI. Also, of course, you should remember the spermatic capacitation.

How do we, as patients judge a clinic based on the embryo laboratory (as we all know the lab is very important) when looking for a clinic. Every clinic says it is the best. Are there any factors we should take into consideration and maybe ask the clinic before choosing the right one? Any factors like the number of IVF cycles per embryologist, experience, certification etc?

It’s difficult to answer this question. When you are choosing a clinic, it’s true every clinic tells you how great and beautiful they are, but when you choose the clinic, you need to choose one with experience, where you feel comfortable and one you can trust. When you feel comfortable, it means the lab is having good practice and it is serious, and once you see the clinic and you meet the team, it is not difficult to detect.

MitoScore—could I ask for your opinion about it? Have you ever used it?

We don’t use MitoScore—we know it, it is an embryo selection technique, but we use time lapsing using an embryoscope to help us choose the best one based on the needs of the patient and we are having very good results, so we don’t think we can improve the results.

There are a lot of rumours about all add-on procedures in IVF like PICSI, AH, EmbryoGlue and more—that they are giving false hope instead of a real treatment option (as I’ve seen on one of the HFEA presentations last year) so do you monitor in some way how much, in terms of the treatment quality, we can get using them- I mean the difference in success rates with or without them?

You are right—there are a lot of techniques and there are places where the techniques are always included in the treatment. As I said before, we only use the ones that we think are useful and the ones that can improve the rates and only when medically indicated and necessary. There are some techniques that are easier to be measured and others that are more difficult to be measured, but results may vary with the techniques, but the results mostly vary when you apply them when necessary.

Do you have any standard procedure (like a set of diagnostics, procedures in IVF Lab etc) for patients with recurrent miscarriages?

Yes, of course, you should have a protocol for recurrent miscarriages. There are some basic things we need to check, and if none of these are positive, then we go one step further.

Would you recommend polar body biopsy and in which cases?

We feel much more comfortable with a blastomere biopsy or a trophectoderm biopsy because these are the ones which have more of a guarantee. We select them depending on the evolution of the embryo because the polar body biopsy has controversial results, so nowadays it’s not clear if it helps or not.

For recurrent implantation failure, would you say PGS is the most important add on? I’ve had 6 embryos fail to implant over 3 cycles and I have 5 frozen embryos left. I’m 38.

PGS is a guarantee before the pregnancy—it is the last step we can perform before the embryo transfer—if you have 5 frozen embryos left, then this is what you should have.

My doctor said it’s not standard practice, but that PGS can be done on frozen embryos—do you agree?

Yes, it can be done on frozen embryos.

There was a mix up before my transfer where my choice to use the EmbryoGlue was not confirmed, so the embryologist said that if I wanted to use it, the embryo would have to sit in the glue for an hour and a half. I asked what she thought if I should do it. She said it would be good for the embryo for it to wait that amount of time anyway. What did she mean by that? I also chose to do assisted hatching. My clinic (IVF Spain) does not believe in EmbryoGlue, but the research I’ve done (and what you have mentioned) shows otherwise.

We believe in EmbryoGlue and we have been using it for years—as we told you, there is a lot of medical evidence supporting that it helps implantation. We have no idea about using it for an hour and a half, as in our practice we normally leave the embryo in it for around 10 minutes before the embryo transfer, and that’s enough to produce good results. Regarding assisted hatching, as we explained before, we don’t use it as a standard, only when necessary depending on the morphology of the embryo.

Can a scratch cause issues to the lining of the womb at a later stage?

Not at all, because we do it before the preparation cycle, so not at all.

If we go with PGS, is the biopsy done on the 5th day or the 3rd day? If it’s 5th day, are the embryos frozen?

We can do both—it is the decision of the embryologists and the geneticists. PGS can be done on day 3 or day 5 depending on the evolution of the embryos and the opinion of the professionals. Doing the biopsy on day 5 does not mean you have to freeze the embryos—it depends on how long the laboratory takes to deliver the results, but normally we would calculate it.

I had an ER mapping done in November which showed that I was pre-receptive. If this transfer fails, would I need the mapping done again? I’ve read that it is only valid for several months?

No, it’s only for that cycle—normally, you should have it done again if you will be doing another transfer.

In your own opinion, if a couple or a woman has only one blastocyst, would you recommend PGD-A?

When you do PGD-A, one of the main reasons is for it to help with selecting which embryo to transfer. If you have one, then the need for selection is automatically gone. There are many factors regarding the couple, for example, medical history and the economic situation, as this is not a cheap technique. It’s a difficult question to answer—we would say it depends on the patient and on the case. To observe the embryos morphologically and a medical evaluation would help to decide.

How long do we usually wait for the results from the laboratory for PGS?

It depends on the study we need to perform, but usually, we have the results in one day.

What do you think is the best progesterone support? When I use pessaries, my period starts on day 8 if implantation fails, but if I use Lubion, my period starts later.

Medication is a very personal thing—it depends on your case, on your levels of hormones. We use different brands vaginally, we use injections… There are some standard protocols, but not all patients are receptive to these protocols, so we would need to know a little more about your history to be able to advise you.

Is ‘ER mapping’ endometrium receptivity testing? Is this the same thing? I had this done as a dummy cycle to determine when it is best to transfer embryos. This happened in November and I am due for my first transfer this February. Are you saying that the testing that I had done will not be valid for a future transfer?

The test that you had done can orientate you, but the problem is that not all cycles for a woman are the same. That’s why we recommend you have it done when you are going to be transferring the embryos.
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Yadira Pallas Galvez, MSc

Yadira Pallas Galvez, MSc

Yadira Pallás Gálvez is a Senior Clinical Embryologist at the Reproduction Unit of Hospital Clínica Vistahermosa in Alicante. She has a degree in Biology from the University of Alicante, Technical and Scientific Research Diploma in the field of Immunology and Histology at the Universidad François Rabelais de Tours, France, and a completed University Specialist Course on Immunohematology at the Universidad Miguel Hernández de Elche, Alicante. Gálvez has been awarded by the Association for the Study of Reproduction Biology (ASEBIR) and has presented numerous publications and talks at Congresses on Assisted Reproduction Technology. She is a member of the Spanish Association of Biologists (COB) and Association for the Study of Reproduction Biology (ASEBIR).