Monday, March 8, 2010

“Immature, Mature and Post-Mature Eggs” - Confusing and Misleading Terminology

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About 38-42 hours following the onset of the spontaneous LH surge in normally ovulating women as well as after the administration of human chorionic gonadotropin (hCG) to women undergoing ovarian stimulation with fertility drugs, the 46 chromosomes in the human egg begin to segregate...to divide in two. This process, known as meiosis or maturational division is designed to leave the mature egg (MII) with precisely 23 chromosomes in its egg nucleus. The remaining 23 are expelled from the egg nucleus (enveloped on a thin membrane) and come to lie in the viteline space (located between the outer membrane of the egg (the oolema) and the egg’s outer shell (envelopment) or zona pellucida. The purpose of the expulsion of this so called first polar body (PB-1) is to ensure that following fertilization by a mature spematazoon (whose chromosome number has also been reduced from 46 to 23), the resulting embryo so propagated would have regain the full quota of 46 chromosomes that makes up the human genome.

Thus the detection by microscopy, of a PB-1 situated immediately under the zona pellucida, indicates that maturational division (meiosis) has been completed. However, it does NOT confirm that chromosome segregation has take place evenly, i.e. that precisely 23 chromosomes remain in the egg nucleus (i.e. that the egg is "euploid"). The presence in the MII egg of one or more chromosomes above or below 23 in number, is referred to as "aneuploidy", a condition that almost always inevitably in failed embryo development, failed implantation, miscarriage or a chromosomal birth defect such as Down's syndrome.

As it turns out, even in younger women a half to one two thirds of MII eggs are aneuploid and this incidence increases rapidly with advancement in age beyond 35 years.

It follows therefore that the absence of a PB-1 on microscopic evaluation (i.e. an MI egg) clearly indicates that the egg had not completed meiosis, and thus "is aneuploid" and “incompetent” (i.e. is incapable of propagating a viable, healthy embryo. On the other hand, the detection of a PB-1 under the zona pellucida (an MII egg) while confirming that meiosis has been completed, offers no assurance that the egg is in fact "euploid" and thus is potentially "competent". Quite to the contrary...most MII eggs are in fact "aneuploid" and thus totally 'incompetent".
It is by and large the chromosomal integrity of the egg, rather than the sperm that determines embryo “competency”. Thus egg “competency" is an essential prerequisite for the propagation of a viable embryo and a healthy baby.

Another interesting fact is that in more than 90% of cases, an embryo that fails to reach the blastocyst stage (>100 cells), will be aneuploid, “incompetent" and thus are doomed from the get go. On the other hand, although not invariably the case, embryos that do make it to the blastocyst stage are much more likely to be euploid and thus “competent.” In fact, even in young women, at least 50% of blastocysts are aneuploid and thus “incompetent.” This percentage increases progressively with advancing age. While age is the main determinant of what percentage of eggs are aneuploid, the protocol used for ovarian stimulation as well as the timing of the hCG “trigger” can also influence the incidence of chromosomal abnormalities. When the hCG trigger is administered too early or too late, the egg might not be developmentally positioned to undergo orderly meiosis and either 1) be unable to expel half its chromosomes as a PB-1 and thus remain an M-I egg, or 2) the PB-1 may be expelled, but could contain an irregular number of chromosomes. In the latter case, the egg would have a visible PB-1 and would thus be labeled as a “mature” (MII) egg, though it would be aneuploid and “incompetent.” The terms “immature” and “post-mature” as applied to eggs , are often interpreted as meaning that the eggs were either harvested (retrieved) too early or too late, and that performing the egg retrieval a day or two earlier or later would have prevented this from happening. This infers that an MI egg was harvested before it was developmentally ready to enter meiosis and that egg post-maturity results from waiting too long before the hCG trigger. This inference is completely erroneous. In fact, an M1 egg could just as easily have resulted from delaying the hCG trigger shot, as it could from administering it too early. Likewise a “postmature” egg can result just as readily from administering hCG too early as too late. For these reasons, the terms “immature” and “post-mature,” as applied to eggs, should be supplanted by the term "dysmature" which simply means that the M-1 or M-2 egg in question is maldeveloped, aneuploid and “incompetent”.

Thursday, March 4, 2010

Upcoming Infertility Seminars Where I'll Be Speaking

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SIRM will be hosting a series of free infertility seminars in March and April where I'll be speaking on fertility issues and breakthroughs as well as answering questions with other SIRM physicians. The dates are as follows:

March 25th: Easton, PA
March 27th: Clinton, NJ
March 28th: Dallas, TX
April 14th: Peoria, IL

The link below has details, maps and registration info:

SIRM Infertility Seminars

I look forward to meeting you!

- Geoff Sher

Thursday, February 25, 2010

ICSI associated with increase in Stillbirth Rate? What a Recent Danish Study Means

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A recent Danish study reported a 4-fold increase in still births following IVF with intracytoplasmic sperm injection (ICSI).

In the past, intracytoplasmic sperm injection (ICSI) , an in vitro-fertilization method, has been used predominantly in cases of moderate or severe male factor infertility. More recently many IVF centers have applied ICSI as a preferred method of fertilization in non-male infertility cases, as well. It is well recognized that when ICSI is performed for male infertility there is a definite increase in embryo defects and related miscarriages.

A few years ago, a large study in Sweden (2003) followed by one reported from Egypt (2004) clearly showed that when ICSI is performed for non male factor infertility, IVF outcome is not prejudiced, and the rate of pregnancy loss and birth defects are unaffected.

Since the Danish study does not differentiate between cases where ICSI was done for male factor versus non male factor infertility, it is likely that their finding of an increase in still births might be due to the effect of abnormal sperm on the embryo’s health rather than being due to the ICSI process itself. And… since male factor infertility requires ICSI, there is no avoiding this procedure in such cases anyway.

Monday, February 22, 2010

Embryo Splitting To Increase the Availability of “Competent” Embryos For IVF

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I recently received several inquiries from visitors to our SIRM discussion boards (http://forums.haveababy.com/index.php?showforum=1 ) on the subject of “embryo splitting” to try and improve the opportunities to conceive through IVF. Here is one example of such an inquiry that recently appeared on the SIRM-Las Vegas regional discussion board:

“I've been reading on embryo splitting (for readers: manually splitting embryos in the lab …..in order to increase the number of embryos to transfer/freeze). The AMA seems to not have an issue with it and understands that it could help infertile couples increase their chances: (www.amassn.org/ama/pub/physician-resources/medical-ethics/code-medical-ethics/opinion2145.shtml)”

Against this background, I wish to share the article below with those of you that may be interested in this topic.
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"Spontaneous embryo splitting resulting in monozygotic ("identical") twinning is a well established peculiarity of nature".

There exists a period during early mammalian embryo development when embryos can be separated purposely into halves or even quarters without drastically reducing the probability that each portion of an embryo is able to develop into a fetus. The usefulness of embryo sectioning is realized any time that the number of embryos suitable for transfer is limited.

When mammalian embryos are surgically split (sectioned) prior to converting into a blastocyst, the number of cells and the size of the resulting blastocysts seem to be unaffected. The developmental time clock also remains relatively unaffected. While bisected embryos develop into blastocysts with slightly less than half the number of inner cell mass cells, and slightly more than half the number of cells forming the trophoblast, these alterations do not seem affect the implantation rate of mammalian, non-human embryos. It remains to be discovered whether the same would apply in the case of human embryos.

Several concerns have been raised regarding the production of abnormal offspring and other potential complications that might result from surgically inducing monozygotic twinning through microsurgical embryo splitting (MES). However, it is a fact that in the cattle industry, where MES has been performed successfully for many years, there has to date been not s single report of abnormalities in the offspring.

Microsurgical Embryo Splitting (MES) might ideally be combined with full karyotyping (chromosomal recognition) using comparative genomic hybridization (CGH) on 6-9 cells, day 3 embryo(s) to identify “competent” embryos. This would be followed by MES on day 3 (the cleaved embryo stage of development), or on day 4 (the morula stage). The resulting hemi-embryos so produced would thereupon be cultured for a few days longer to the blastocyst stage when they would either be transferred fresh to the uterus or be vitrified (frozen) and banked for subsequent dispensation. In this manner MES might serve as a means to increase the number of available embryos per woman.

Potential complications such as twin-to-twin transfusion, umbilical cord entanglement and obstructed birth, which often occur with natural spontaneous monozygotic twinning, are not encountered in MES-induced twinning. The explanation lies in the fact that unlike natural monozygotic twins that often occupy a single, common gestational sac and frequently share a common blood supply derived from a single placenta, MES-induced monozygotic twins invariably have their own placentas and reside in separate gestational sacs.

MES, should it become a feasible, acceptable and practicable clinical option in ART could by increasing the number of available viable embryos offer hope for thousands of women who because of advanced reproductive age and/or diminished ovarian reserve are only capable of producing very few "competent" eggs/embryos at a time. It might also prove to be of benefit in cases where (out of personal choice) IVF is performed without the use of fertility drugs (Natural Cycle IVF) and where accordingly only one or two eggs/embryos are generated. ----------------------------------------------------------------------
Addendum: SIRM-Las Vegas is soon to launch a limited study to evaluate the efficacy of MES. Criteria for consideration of inclusion in this study will be as follows:

• Age less than 36 years • Regular menstruation/ovulation
• Normal endometrial cavity
• Absence of a significant male factor infertility.
We intend, provisionally to focus this study on young couples considering Natural Cycle IVF.
Please email Tom Anderson (
toma@sherinstitute.com) if you would like to be considered for inclusion in this proposed study.

Tuesday, February 16, 2010

Egg Quality and Ovarian Reserve: The Effect of FSH and Inhibin Blood Levels

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In most cases, when basal blood inhibin levels fall, the woman’s Follicle Stimulating Hormone (FSH) level will rise. It is likely that inhibin produced by the granulosa cells lining the cavity of early follicles signal an area in the brain known as the hypothalamus which produces small “releasing hormones” that direct the woman’s pituitary gland to regulate the production and release of FSH. This also explains why basal inhibin and FSH as well as the number of early follicles (antral follicles) represent reasonable measures of ovarian reserve, which in turn correlates with the woman’s potential to produce follicles on her own and/or in response to ovarian stimulation with fertility drugs.

High FSH and low inhibin/antral follicle count (AFC) in the first few days of a woman’s cycle suggests diminished ovarian reserve and serve as a warning that the woman will not likely produce an optimal number of mature follicles and eggs in response to ovarian stimulation. They also serve to encourage the use of a more aggressive, yet individualized approach to ovarian stimulation.

Most women develop diminished ovarian reserve about 6-8 years prior to the onset of menopause (that time the period is termed the “climacteric”) but in some cases this happens at a much younger age (i.e. “a premature climacteric”). Thus, the basal FSH, inhibin level, and AFC are quite good indicators of ovarian reserve and the number of follicles that are likely to develop, given an optimal protocol for ovarian stimulation. However, these parameters alone are not good predictors of subsequent egg and/or embryo quality. Rather it is the woman’s age and the protocol designed to effect ovarian stimulation that play the major role and here, these measurements of FSH/inhibin and AFC can assist in designing the ideal stimulation protocol.

Let me explain! Human eggs undergo degradation in quality over time, such that by age 39, an egg (ovulated or harvested at egg retrieval) will on average have about a 20% chance of being genetically/chromosomally normal. This is about one half the chance at age 35 and under. By the time she reaches her mid forties, that number will decrease by half again (i.e. reaching less than 10%). This "wear and tear" effect on egg quality is an inevitable consequence of the advancing "biological clock". So, when it comes to egg quality, it is the woman's age and the protocol of ovarian stimulation that are the most important determinants. You simply cannot stimulate a woman in her 40's or for that matter a woman with diminished ovarian reserve using the same "recipe" (i.e. the stimulation protocol) as you would prescribe for a younger woman who has normal ovarian reserve. If you do not individualize the protocol of stimulation, you are highly likely to propagate the development of poor quality eggs that have a disproportionately increased likelihood of having chromosomal abnormalities.

Again..... the most important factors affecting a woman’s egg quality are 1) her age, and 2) ovarian reserve. While these two variables may be linked (women are more likely to develop diminished ovarian reserve as they get older), women sometimes do experience a “premature climacteric” and egg quality deteriorates with advancing age regardless of ovarian reserve. Thus, these two contributing factors should be seen as related, but independent variables.

So how then does age and/or diminishing ovarian reserve affect egg quality? First, as stated above, it is inevitable that with advancing age, egg quality will decline. Second, for follicles to grow and for eggs to develop normally (an essential prerequisite for proper genetic maturation), the tissue surrounding the follicle (ovarian stroma or theca) must produce testosterone. Stromal testosterone production requires luteinizing hormone (LH), which is produced by the woman's pituitary gland and is also acquired through some varieties of injectable fertility medications (Menopur, Repronex and Luveris) given in the course of ovarian stimulation. The testosterone then gets carried in "bucket brigade” fashion to the granulosa cells lining the inside of the adjacent follicle(s). Here, under the influence of FSH, it gets converted to estrogen (mainly estradiol/E2). In the process, the follicle grows and the egg it harbors within undergoes vital developmental changes in preparation for final genetic maturation ("ripening") that occurs with the spontaneous surge of LH that triggers ovulation (or following the administration of the hCG during ovarian stimulation).

Thus, a small amount of testosterone is needed for optimal egg quality (though too much testosterone can be harmful to the egg as I will discuss below). Eggs that have the genetic potential to transform into genetically “competent,” mature eggs will do so within 36 hours of the spontaneous pre-ovulatory LH surge, or following hCG-induced ovulation.

The important consideration here is that there should not be preovulatory over-exposure of the developing egg to testosterone, something that is most likely to happen when older women and/or those with premature diminution in ovarian reserve are prescribed a suboptimal protocol for ovarian stimulation. If there is overexposure to testosterone, egg development and subsequent egg/embryo quality can be severely compromised along with the chance of a healthy pregnancy. Since older women (>39 years) and women with diminished ovarian reserve tend to produce an excess of LH and have a tendency to over-produce testosterone, this is where the problem lies.

What does this all mean in the context of preparing a woman for a cycle of ovarian stimulation? First, it means that a young woman who has diminished ovarian reserve should still be capable of producing produce good quality eggs, albeit in a smaller number, provided that she gets prescribed an individualized and customized protocol that is designed to prevent over-production of ovarian testosterone. Conversely, an older woman with diminished ovarian reserve will, because of the inevitable effect of age on egg quality, produce a higher percentage of poor quality, genetically “incompetent” eggs.

Finally, when it comes to natural cycle IVF, it should be recognized that some women will produce up to 2 or 3 follicles. Some will be smaller than others, but even the smaller ones can yield eggs. However, as with regular IVF (with ovarian stimulation), the quality of her eggs will be inevitably be influenced by her age. Thus the success rate following natural cycle IVF in such cases will be very much lower than for younger women, especially if they have diminished ovarian reserve. The success rate with natural cycle IVF, even in young women, is only about 10% per cycle. In older women, it is under 5%. Furthermore, older women are more likely to have increased LH production. In addition, the LH they produce becomes more potent with advancing age and thus is much more likely to evoke a greater ovarian testosterone response with a negative effects on egg/embryo quality (and correspondingly on the likelihood of IVF success).

In summary, natural cycle IVF is a much less effective and successful form of IVF across the age spectrum. If it is used, it should be confined to younger, ovulating women who have a normal ovarian reserve.

Wednesday, February 10, 2010

The “Vanishing Twin”: What Does It Mean?

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Today, in first world environments where there is ready access to advanced medical technology, many women undergo ultrasound diagnosis of pregnancy as early as 5-6 weeks after their last menstrual period. As a result, multiple pregnancies are usually recognized very early on. Serial ultrasound follow-up examinations in such cases have shown that often times one of the developing babies “mysteriously” vanishes, with the remaining conceptus (baby) usually proceeding to a healthy birth. Since most multiples comprise twin pregnancies, the term “vanishing twin” is used to describe this situation. While in most cases the loss of a vanishing twin is associated with painless, innocuous bleeding, this is not always the case. In fact bleeding might not occur at all.

The incidence of spontaneous pregnancies resulting in twin births is about 1:80. In women under 35 treated with clomiphene citrate it is about 1:20. After ovarian stimulation using injectible gonadotropin fertility medications, it increases to approximately 1:5 and after IVF (given the current tendency to replace multiple embryos), the incidence of twins is about 1:3.

What many fail to appreciate is that about 1 in 10 spontaneous pregnancies start off as twins, but as the pregnancy advances into the 1st trimester and beyond, one twin will “vanish” (absorb) while the other will continue as a healthy, unaffected singleton. When this happens, the occurence of painless mild bleeding (or spotting) raises understandable concerns by the affected woman who often asks:

Q.Am I about to miscarry?

Answ: The bleeding results from the absorption of one of the pregnancies, and since the vast majority of twin pregnancies have separate and independent placentas, the loss of one will accordingly usually not affect the remaining twin. As long as the bleeding remains mild and the woman does not experience an increase in cramping and pain over a period of a few days, the pregnancy will probably not be lost. In fact, in the majority of such cases this is precisely what happens.

Q. How long will I continue to bleed?

Answ: In most cases - unless the pregnancy is destined to miscarry completely - the bleeding will remain painless, mild, and will stop within a week or so. However, the timing will depend on when the conceptus was lost. If this occurred late in the first trimester, the bleeding will usually last longer (even a few weeks) than in cases where the pregnancy was lost earlier. It should be that in some cases , and the body reabsorbs one twin with no outward indication (bleeding) of the loss.

Q. How will the loss of one twin affect the surviving one?

Answ: In the majority of cases, the remaining conceptus will progress unaffected, to a healthy birth.

Q. - Will there be any remaining evidence of the vanished twin at birth?

Answ: Usually not! Sometimes a small area of scarring or “thickening" of part of the placenta will be seen at birth. However, this usually only occurs in cases where the first twin succumbed late in the first trimester or beyond (in the 2nd trimester).

Q.Could the vanishing of one twin have been prevented? Did I do something wrong?

Answ: The vanishing twin is not the result of something the mother/father did - or failed to do. In most cases, the vanishing twin is lost for the same reason that single pregnancies miscarry (i.e. because af chromosoma aneuploidy). In a small number of cases the loss results from autoimmune or alloimmune implantation dysfunction. Since many vanishing twins are lost very early in pregnancy, before most women will have undergone an ultrasound to confirm pregnancy, most cases go undetected. The woman would have no knowledge that she had been carrying more than one fetus. In fact, as stated above, many more of us begin life as twins than was previously thought.

Vanishing concepti can also occur within high order multiple pregnancies (triplets or greater). A triplet pregnancy can reduce spontaneously to a twin or singleton and so can a quadruplet pregnancy.

In the final analysis, individuals and families who experience the "vanishing" of a conceptus will experience anxiety and even panic when bleeding starts, followed by a sense of relief when it finally stops, and they learn that the remaining fetus and the pregnancy have survived. This will in most such cases be followed by a profound sense of loss and sadness , especially if the woman and/or her partner had been looking forward to a twin birth.

Thursday, February 4, 2010

Share Your Stories on the SIRM Facebook Page

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Many of our patients have shared their inspirational stories on our Facebook page. We would love to hear yours! Just follow this link:

http://www.facebook.com/pages/Sher-Institutes/213018641760