Culture of embryo's
14 Replies
4everhopeful - April 20

Dear Dr Smith,

I recently found out by chance, after developing swellings in my joints, post IVF failure, that I have low titre ANA 1:80. When I told my RE she said it could account for why we haven't had any success so far...

She referred me for further tests at Millenova in Chicago, and it turns out that I have DNA problems with MTHFR (heterozygous), Factor X111 (heterozygous) and PAI-1 (homozygous).

I was told that our embryo's will now need to be cultured in a specific way, using hyaluron acid G3 series, which apparently no clinic in the UK (where I live) does :o and that we'll now have to travel to the USA for tx.

I was wondering if you can shed any more light on what this culturing of embryo's is, and why we have to have it, if we're to stand any chance of success?

Many many thanks,

Nat :)


Dr Smith - April 21

I have never heard of clotting abnormalities mandating specific embryo culture conditions, so its wierd to me too.

The G series media (G-Fert, G-1, G-2) are made by Vitrolife and are FDA-approved (US) for embryo culture (I personally use this media for IVF). All cell culture media require supplementation with protein. Usually, human serum albumin (HSA, derived from human blood) is used. Vitrolife also has recombinant serum albumin (rHSA, manufactured by yeast) The G-series media may be supplemented with either source of protein. Recombinant hyaluronic acid (rHA) is present in Vitrolife's "Embryo Glue" product used for embryo transfer.

I'm thinking, because of the clotting disorders, that your program may be sugesting that you have your embryos cultured with all recombinant products (rHSA and rHA) to avoid any contact with human blood products. I don't see how this could make a difference, but they may know something I don't.

Clotting disorders are usually treated with low molecular weight heparin.


4everhopeful - April 21

Thank you so much for your prompt reply Dr Smith :)

My clinic in the UK were initially hopeful that they could help me and obtain the culture medium we needed, but when the embryologist contacted Chicago, he was told that it was a specific process (something to do with spinning?) that we needed, and that this process isn't available anywhere in the UK at the moment :-\
Do you think it must be a really new technique, as it seems weird to you to? But then you would still have heard of it... :-\ I'm so confused!

To combat the effects of the clotting disorders I've been advised to go on metformin and take extra folic acid (800mcg plus 1mg), though I've heard others with the same problem are taking Folgard, which isn't available in the UK :-\

Do you treat clotting disorders with heparin then at your clinic? Only nothing was mentioned to me about taking blood thinners :-\

A lady I "talk to" on-line, who has similar DNA problems as me (who is undergoing IVF tx at a clinic in New York), says she has to take clexane.
What's the difference between heparin and clexane?

I'm very confused about all this, and still dumb founded that no one in the UK can help us ???

Many thanks for all your help :)

Nat :)


Dr Smith - April 22

I think I better defer to Dr Jacob at this point. He would be more familiar with the drug treatments for these kind of clotting disorders.

I'm also confused by what the lab is telling you. This is all news to me. I'm afraid I can't comment, since I haven't a clue what they are talking about. Sorry.


4everhopeful - April 25

Thank you for all your help Dr Smith, I'm very grateful to you :)

I'll take your advice and post this to Dr Jacob to see if he can shed any light.

With warmest regards,

Nat :)


4everhopeful - April 28

Dear Dr Smith,

I followed your advice, and Dr Jacobs wasn't aware of it either and referred me back to you!!

Anyway, I thought you'd be interested to know that I had a phone consultation with my RE in the UK today, who has been in contact with the clinic in Chicago who said we needed specific culturing of our embryo's... Apparently it's based on the research of one embryologist and is still only in the experimental stages, which is why she said it's not available or known about in the UK or US.

Being just an ordinary lay person, I don't understand the process at all, but my RE said something about enzymes in Hyaluron Acid G3 series being benefitial to implantation :-\ ??? Does that make any sense to you? I found a study on the internet about mice with mutated PAI-I and how specific culturing of their embryo's resulted in improved implantation rates (75%) but found no similar studies for their human counterparts!

Because neither you nor Dr Jacobs were aware of it, it's prompted me to question why I had no choice but to go to the US for tx :-\ Now that I know it's only experimental, I'm not sure I want to risk us having to spend more than double for something which is still largely unproven.
I'm always though grateful to any new research, and we haven't ruled out trying this in the future if we go on to suffer more IVF failures, but are hoping to get lucky here first and not need it!

So a big thank you for all the help you have given me :)

With warmest regards,

Nat :)


Dr Smith - April 28

Ahh, the lightbulb above my head is now dimly lit. I think they may have been talking about the use of hyaluronan (a long-chain, naturally ocurring, polysaacharide) added to G-2 medium for embryo transfer. There is a commercially available product in the US called "Embryo Glue", manufactured by VitroLife. At least in VitroLife's sponsored studies, Embryo Glue was shown to have a benificial effect on implantation. This may have been due to a molecular bridging effect between the embryo and the endometrium facilitated by hyalronan or hyaluronan's ability to increase the viscosity of the transfer medium (the embryos stay where you put them after transfer). There have been no independent, large scale, clinical studies to support this notion. However, I've been using Embryo Glue as a blastocyst transfer medium for about 3 years and we saw a small, but significant, increase in implantation rate after we started using it. We never published these findings because I do not believe in promoting any particular product.

HOWEVER, using Embryo Glue to treat clotting disorders is completely uncharted waters.


4everhopeful - April 28

Thank you so much for the info on "Embryo Glue" Dr Smith :)

I will print this page off and show it to my RE, because she seems very open to trying different approaches.

I appreciate that it's benefit for treating clotting disorders is unproven as yet, but I think I'd really like to try this next cycle if it's possible :-\ Unfortunately my clinic doesn't do blastocyst transfer, and only ever trasfer embryo's on day 2, so would it still be possible to use this to culture our embryo's?

Many thanks for all your help :)

Nat :)


Dr Smith - May 2

"Embryo Glue" was designed as a embryo transfer medium. The composition of the medium is appropriate for the nutritional requirements of embryos that have reached the 8-cell stage or beyond (i.e. morula, blastocyst). It was not designed for embryo culture from Day 1-2 and may compromise development. I wouldn't recommend it. Sorry.


4everhopeful - May 4

Thank you for letting me that know Dr Smith.

It seems I would have to request a later transfer than day 2, and the embryologist at my clinic is very anti blastocyst transfer :-\ but being headstrong I will still request it anyway!

Am I right in my understanding that you use a different culture up to day 3 and then transfer them to embryo glue for their continued development?

Many thans for all your help :)

With warmest regards,

Nat :)


Dr Smith - May 4

In a sequential media culture system, one medium formulation is used for fertilization (Day 0-1), another media formulation for Days 2-3, a third media formulation for Days 3-5(or 6) and then a final media containing hyaluronan (Embryo Glue) for the transfer of the blastocyst stage embryo. These media formulations are designed to meet the stage-specific nutritional requirements of the sperm, eggs and embryos throughout development to the blastocyst stage. The formulations are based on years of scientific research directed toward understanding human sperm, egg and embryo metabolism. Thankfully, gone are the days of "guess work". However, as research in this field proceeds at break-neck speed, these are formulations are revised regularly as new information become available.

An embryologist that is vehemently anti-blastocyst is not a good sign. Many programs (in the US at least) culture embryos to the blastocyst stage for cryopreservation, even if they transfer 2-3 of the best embryos on Day 3. An unusually strong anti-blastocyst position may reflect the use of an outdated culture system or, at the minimum, outdated thinking. Blastocyst culture technology was originally developed in Europe in the early 90's, so this is not just an "American fad". Are you reasonably sure about your chances of success at this program? I'm beginning to worry...


4everhopeful - May 4

Thank you for getting back to me so quickly Dr Smith :)

The clinic I cycle with used to offer blastocyst (for an extra fee) but it was then stopped, because the embryologist who is there now, is of the opinion that they're better off inside, so blastocyst transfer is no longer an option. Though in the UK more people than not, have e/t on day 2.

The clinic I'm at does have a sister hospital in London which offers blastocyst, so we could go there instead :-\ I've always been interested in it, but have always been talked out of it in case 'there's nothing left to transfer'.

Last cycle we had 8 embryo's. 2 were grade 1.5 (1 being best at this clinic) 5 cells on day 2, and the rest were all lower grades. I had the best 2 transfered, but the rest had all arrested and died by the time they would have reached the blastocyst stage :( But then again I've no idea what they were cultured in, esp seeing as the clinic don't do blastocyst culture :-\ They were going to destroy them on day 2, but then decided to give them a chance over the weekend to see if we could have any for freezing.

In light of what we know about how specfic culturing of our embryo's 'may' help them to stick with my blood clotting problems, would I be best to find a programme that offers us blastocyst culture?
Do you think in light of what happened to our left over embryo's last time, we would stand a chance of getting at least 2 to the blastocyst stage in the lab? Do you think it all depends on the conditions of the lab too?
In your programme is it usual to get at least 2 to the blastocsyst stage?

Sorry for asking so many questions, but I really want to explore all the options and give it the best shot we possibly can.

Many, many thanks for all your advice, I really appreciate it.

With warmest regards,

Nat :)


Dr Smith - May 4

When culturing to the blastocyst stage, embryo culture conditions do play a part. In the US where sequential culture systems are readily available and emplyed, it is not a big issue. The G-series media manufactured by Vitrolife, including Embryo Glue, is distributed in the UK through:

Research Instruments Ltd.
Bickland Industrial Park
Cornwall TR11 4TA
United Kingdom

Contact: Ms Kirsty Wilkinson
Tel: +44-1326-372 753
Fax: +44-1326-378 783

Other European media manufacturers also have sequential media systems, Vitrolife doesn't have a corner on the market.

I would contact the program(me ;)) in London to find out what culture system they use for blastcysts and if they routinely culture extra embryos to the blastocyst stage for freezing. If so, I'd say you're in good hands. I would arrange a consultation with a doctor in the London program to get their take on your situation. In someone your age, under the appropriate culture conditions, I'd expect approximately half of the embryos to make it the blastocyst stage. If you get 8 eggs, then two blastocyst stage embryos is a reasonable expectation.


4everhopeful - May 5

Thank you so much again for getting back to me so quickly Dr Smith :)

My dh and I hae been very confused by all this of late, I've even talked to him about the possiblities of coming to your clinic... but the down side of us having to travel to the US for tx, is cost unfortunately :-\
But in the meantime we will definately contact the sister clinic in London to see what they suggest for us...
If we could get 2 to blast I'd be very happy indeed!!

Because I have these genetic mutations though, would that make my eggs more likely to have chromosone abnormalities?

Many many thanks for all your help and advice,

With warmest regards,

Nat :)


Dr Smith - May 5

I am not a geneticist, so I'm talking out of turn here. I'll tell what I know, but for a real, scientific answer, perhaps you should consult a genetic counsellor. We have successfully treated individuals with your clotting disorder mutations. They are not that uncommon in the repeat IVF failure group. When I reviewed the embryology reports for these cases, I did not see anything abnormal about the embryo development (i.e. normal blastocyst development for their age). So, as far as I can tell, there is no link between these mutations and embryo development per se. The impact of these mutations appears to be entirely post-implantation and from the uterine side of the equation.



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