Key Lectures

100 Pages·2015·1.71 MB·English

Abstracts of the 6th Yazd International Congress and Student Award in Reproductive Medicine with 1st Reproductive Genetic Congress
Key Lectures on predictor sheets to discuss with the immunologist,
embryologist and genetic specialist to design a protocol

for induction and embryo freezing. Based on the results,
K-1 this integrated approach can significantly improve the
Past present and future of human reproduction IVF cycle outcome thus it should be offered to the
patients to achieve the best possible results.
Frydman RFL.
Department of Obstetrics and Gynecology, Hospital Foch de
K-3
Suresnes, Suresnes, France.
Surgical or medical treatment for unruptured
Email: [email protected]
interstitial (cornual) ectopic pregnancy? That is
Since the last forty years, Assisted Reproduction the question
Technologies (ART) open a new area for infertile
couple. Most of the etiologies of sterility such as tubal Rahimi M.
Department of Obstetrics and Gynaecology, Grey Base
blockage, ovulation perturbation or endometriosis can
Hospital, Greymouth, New Zealand.
be treated. Ovarian stimulation, in vitro fertilization,
Email: [email protected]
intra cytoplasmic injection and freezing approach is
now available for both female and male factors. But two Cornual pregnancy is a rare and most dangerous form of
black boxes have to be explored: the uterus capacity to ectopic pregnancy (EP) which is usually treated by
implant and the capacity of each gametes and embryos corneal excision or hysterectomy. The consequence of
to develop. Genetic, epigenetic and immunological corneal location of gestation is usually massive
approaches will be the next steps of knowledge in order intraperitoneal haemorrhage, necessitating a blood
to increase the results and to give a personalized transfusion. Controversies exist between the group of
proposition for each couple. gynaecologists who excise the corneum via laparoscopy
or laparotomy and the group of gynaecologists who
K-2
leave the corneum intact and use drugs (i.e.
The improvement of IVF cycles outcome: A new Methotrexate) for treatment of corneal EP. Expectant
approach management of this type of EP is suitable only for
women with low and diminishing levels of βHCG.
Nemati M1, Nemati Sh2, Rafeizadeh Gh1. Each group claim their way of managing unruptured EP
1. Shahrekord University of Medical Sciences, Shahrekord,
is preferable over the other method. There are
Iran.
advantages and disadvantages in each mode of
2. Shahid Beheshti University of Medical Sciences, Tehran,
Iran. treatment. Patients with corneal EP usually have signs
Email: [email protected] and symptoms of ectopic gestation later in the first

trimester of pregnancy. This is because the location of
Integration of basic science and clinical study of
the gestation allows more room for the growing EP.
infertile patients result in improving the success rate of
Therefore the size and the level of βHCG are higher
IVF cycles. Unfamiliarity and or lack correlation
than other types of EP. In fact there are anecdotal
between basic and clinical scientists involved in
reports of term interstitial EP. In view of this fact, these
reproductive sciences lead to ignore some reproductive
patients with high levels of βHCG are not suitable for
system disturbances that can affect ART cycles
medical therapy. Currently laparoscopic surgery is the
outcome. According to financial and emotional burden
preferred treatment for EP. There are 2 laparoscopic
of the failure of ART cycles, it seems that the time has
techniques:
come to modify ART from relatively the same form of
1. Laparoscopic wedge resection of the corneum which
drug administration for all patients or exam and error on
involves removal of the myometrium surrounding
them to design the specific ART cycles based on the
the interstitial section of the tube. This results in
initial characteristics of the patient‟s reproductive
higher risks of uterine rupture in the subsequent
system. In this approach, IVF/ET protocols being
pregnancies.
designed only after close monitoring of each patient‟s
2. A simple, swift and safe (SSS) laparoscopic
natural cycle to identify; the initial characteristics and
technique for the treatment of interstitial pregnancy
disturbances, antral follicular count, follicular and luteal
is applying 2-3 vicryl endoloops below the affected
phase length, the patient‟s endocrine profile, the largest
corneum incorporating the proximal third of the
size of dominant follicle, partial or complete rupture of
tube, mesosalpinx and portion of the myometrium
dominant follicle, grading of the endometer, occurrence
adjacent to the corneal EP. This should be done after
of premature luteinized or delayed maturity of
cornuostomy and suction evacuation of the products
endometrium, the characteristics of previous induction
of conception in the corneum. This technique is easy
cycles and the probability of initial or final oocyte
to perform by any gynecologist whom has some
atresia. All of these factors need to evaluate and record
experience in laparoscopy.
Iranian Journal of Reproductive Medicine, Vol. 13, No 4 (Suppl. 1), April 2015 1
Abstracts of the 6th Yazd International Congress and Student Award in Reproductive Medicine with 1st Reproductive Genetic Congress
This technique was developed by the author in New Fatemi HM.
Zealand in 1995 on an unexpected corneal EP Nova-IVI, Abu Dhabi, UAE.
Email: [email protected]
undergoing emergency laparoscopy. This case was

presented at the 26th Annual Meeting of the American
Infertility is worldwide acknowledged as a major health
Association of Gynaecologic Laparoscopists in Seattle
concern. Although infertility prevalence appears to
(September 23-28, 1997) and published in the Journal of
remain unchanged since the 1990s, significant regional
the American Association of Gynaecologic
differences have been reported in infertility prevalence.
Laparoscopist (May 1999, Vol. 6, No. 2) as a new
The prevalence of infertility in women of reproductive
laparoscopic approach for the treatment of interstitial
age has been estimated to be one in every seven couples
ectopic pregnancy. Following this laparoscopic
in the western world and one in every four couples in
treatment the patient had 3 more pregnancies which
developing countries. Geographical, sociocultural/
were all intrauterine and in the last pregnancy the baby
religious and ethnical dissimilarities contribute to these
was born by caesarean and tubal ligation was performed
global variations of infertility prevalence. Infertility has
as per patient request and consent in 2001. In the
a major impact on family stability in many cultures,
author‟s opinion the above laparoscopic technique is
especially in developing countries, where childlessness
preferred to the medical treatment because it ends the
can impact sociocultural status. Moreover, it is
EP and its risks and patients anxiety in one session.
important to realize that most fertility treatments are
Methotrexate may be a reasonable option in selected
based on studies performed in Western countries. The
women with a low βHCG level but is not successful in
purpose of this review is to critically appraise the
every interstitial pregnancy.
existing evidence regarding the association between

female fertility and relevant sociocultural factors in
K-4
Middle East countries focusing on aspects such as
Ultrarapid freezing “Vitrification” is the right
parental consanguinity, obesity and vitamin D
tool for cancellation of fresh embryo transfer
deficiency. There may be reason to believe that in

addition to the current standard evaluation of infertile
Al-Hasani S.
couples, region-specific counselling and treatment
Reproductive Medicine Unit, University of Schleswig-Holstein
modalities are required.
at Luebeck, Luebeck, Germany.

Email: [email protected]
K-6
Single embryo transfer is becoming increasingly Ultrasonography screening in obstetrics in
popular in IVF/ ICSI. More IVF/ ICSI cycles therefore perinatal medicine. What, when and by who?
include freezing of high quality embryos, and the
cumulative effect of such cycles becomes more Antsaklis AJ.
important. To improve the results obtained using frozen- Division of Maternal Fetal Medicine, Department of
thawed embryos, the predictive value of embryo and Obstetrics and Gynecology, Alexandra Hospital, School of
Medicine, University of Athens, Greece.
patient characteristics such as ovarian reserve, hormone
Email: [email protected]
levels and age play an important role in both cases

whether the women treated with Oestradiol/
Ultrasonography represents the most significant
progesterone or undergo natural cycle transfer.
advance in obstetric diagnosis and clinical management
Although, embryo quality indicators revealed sometime
in the past 40 years. Ultrasonography in pregnancy is a
morphologically and numerically inferior embryo
simple, painless and harmless examination used in
cohorts after cryopreservation, the clinical pregnancy
everyday practice for the present diagnosis. The largest
rate is higher in cycles using thawed embryos compared
risk of antenatal sonography is probably misdiagnosis.
with fresh embryos. Moreover, subsequent logistic
A false positive diagnosis of a malformation may lead
regression analysis controlled for differences in embryo
to parental anxiety and these errors can be corrected by
quality and revealed significantly greater probability of
a second examination in a tertiary referral center. A
clinical pregnancy with thawed embryos when
missed diagnosis (false negative) remains undetected
compared with fresh embryos, suggesting a negative
unless the patients undergoes for a second examination
effect of ovarian stimulation on endometrial receptivity.
for another indication. These limitations are often
The aim of this study is to discuss an idea of
gestational age dependent. But if a significant
cancellation of a fresh embryo transfer and put on an
congenital anomaly is recognized at delivery one of the
alternative method which is the frozen thawed embryo.
patients question is: «Could we have seen this on

ultrasound before delivery?» Obstetrics sonography
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should be performed at an appropriate gestational age
Sociocultural influences on fertility in the
by an experienced practitioner.
Middle East: the role of parental consanguinity,
The ACOG and the AIUM have published guidelines
obesity and vitamin D deficiency
for the basic ultrasound examination in pregnancy.
2 Iranian Journal of Reproductive Medicine, Vol. 13, No 4 (Suppl. 1), April 2015
Abstracts of the 6th Yazd International Congress and Student Award in Reproductive Medicine with 1st Reproductive Genetic Congress
This basic examination is performed most often for the to the use of ultrasound. Limitations of obstetrical
purpose of biometry and the establishment of ultrasonography should be briefly reviewed with
gestational age. Various descriptive terms have been patients prior to the initiation of the procedure. Some
used to identify such a detailed study including level II major malformations are easily detectable whereas
comprehensive, extended and targeted. This targeted other malformations present subtle ultrasound images,
study is performed for the detection of fetal anomalies and may not be diagnosable in the midtrimester.
in women at risk for having a malformed fetus. The Ultrasound is used not only for diagnosis but as a tool
pregnant patient expects to have information about for the management of a complicated pregnancy and
baby's health and in case a congenital anomaly is for this reason the perinatologist is perfectly the right
present she wants to kwon the prognosis, the treatment doctor to provide sonographic diagnosis and plan the
and the recovery. Routine use of ultrasound in low management of a high risk pregnancy.
pregnancies has been offered for the decrease of labor Conclusion: The issue of routine sonography for low
inductions performed for postdatism, for the early risk pregnant women continues to be contentions even
detection of multifetal gestations, for detection of though, randomized trials have not been able to
placental implantation abnormalities and for the demonstrate a clear benefit. Although great progress is
antenatal diagnosis of congenital anomalies. being made in the first trimester diagnoses of
There is good evidence to support the recommendation congenital anomalies, most targeted studies are
that the sensitivity of the ultrasound screening in performed at 18-20 weeks of gestation. The highest
detecting fetal malformations in low risk pregnancies rates of detection of congenital anomalies are seen in
cannot be established with precision it will continue to tertiary care settings such as a university medical
be decided on a local level and varies in different center. In high risk cases a consulting perinatologist is
centers with different level of operators training and commonly the physician most likely to integrate the
financial resources. Sonography for fetal biometry and ultrasound findings.
when precise estimation of gestational age is required
(in cases such as planning a caesarean delivery), should K-7
be performed in the first trimester or as early in The role of hysteroscopy in female infertility
pregnancy as feasible. 18-20 weeks is the traditional management
and appropriate time to perform a targeted scan. This
ultrasound study allows a detailed review of fetal Karimzadeh MA1, 2.
anatomy and is early enough so that amniocentesis or 1. Shahid Sadoughi University of Medical Sciences, Yazd,
Iran.
other diagnostic procedures can be performed prior to
2. Madar Hospital, Yazd, Iran.
fetal viability.
Email: [email protected]
The genetic sonogram is a targeted study with special
emphasis on ultrasonographic markers that may Implantation is an important factor that was influenced
indicate aneuploidy. Targeted ultrasonography at 18- with the embryo and endometrium dialogue. The
20 weeks allows the couple to consider all of their uterine evaluation before any assisted reproductive
options and allows for appropriate referral and technique should do as a routine procedure.
counseling. However some malformations are not Hysterosalpingography is the first method of uterine
easily visualized at this period. Hydrocephalus or abnormality evaluation. But different researches have
bowel atresia‟s may develop after this period and may shown the false negative result of HSG in uterine
not be demonstrable until after 24 week's gestation abnormalities in 18.4%. Hysteroscopy is the gold
while the optimal time for fetal echocardiography is standard procedure for uterine cavity exploration
probably somewhat later (20-22 weeks). through direct visualization in patients with recurrent
By whom: Antenatal sonography is performed in implantation failure. It appears that more than 1/3 of
different medical centers, doctor's offices, hospitals, by the patients interpreted as normal following HSG are
physicians of varying levels of experience or by found to have a uterine abnormality after diagnostic
technicians. If a physician is unable to document hysteroscopy, which might be a significant cause of
formal residency, fellowship, or other postgraduate reproductive failure.
training, he or she must have completed 100 hr of Polyps are the most common pathological lesions in
American Medical Association category 1 continuing infertile women especially in unexplained infertility.
medical education in diagnostic ultrasound, with The possible role of these polyps in infertility is yet
evidence of involvement at least 500 diagnostic unclear but surgical removal of all endometrial polyps
examinations under the supervision of a qualified among infertile women is crucial. Removal of polyps
physician. The experience of the obstetrician clinician may enhance reproductive outcome between 43-80%.
with sonography must begin with detailed knowledge The mullerain abnormalities in the uterus such as
regarding fetal cross sectional anatomy. It is important septate, subseptate, arcuate and bicornate are common
for the clinician to know his or her limits with regard findings in the hysteroscopy of repeated IVF failure
Iranian Journal of Reproductive Medicine, Vol. 13, No 4 (Suppl. 1), April 2015 3
Abstracts of the 6th Yazd International Congress and Student Award in Reproductive Medicine with 1st Reproductive Genetic Congress
patients with previous normal HSG. Although WHO is Sonography was found to have up to 84% sensitivity
recommended the office hysteroscopy when clinical or and 90% specificity for the detection of endometriomas,
complementary exams such as ultrasound and HSG confirmed by surgery and histopathology. The goal of
suggest intrauterine abnormality or after IVF failure surgical treatment is to remove visible areas of
but it is a routine procedure before the first ART cycles endometriosis and restore normal anatomy by lysis of
due to enhancing fertility. Implantation improvement adhesions.
after hysteroscopy could be related not only to treating
uterine cavity lesions but also may be affected by K-9
cervical canal dilation and evaluation of the direction First and second trimester screening for
of the cervical canal for easy embryo transfer, aneuploidy
assessment of interior of the uterine cavity and shape
abnormality. Moreover, the uterine instrumentation Wahaaj MH.
cause endometrial injury and stimulates inflammatory Wahaaj Hospital, Kabul, Afghanistan.
reactions to growth factors and may improve the Email: [email protected]

pregnancy rate by near 2 folds (32-44% vs. 21-26%).
Over the last two decades, risk assessment for
Although the position of hysteroscopy in infertility
aneuploidy has been refined to the point that maternal
management is unclear but diagnostic hysteroscopy is
age alone is no longer considered adequate in
a valuable test and should be advised routinely as part
determining the risk of having a chromosomally
of patients' investigations before IVF/ET.
abnormal offspring. Obstetric sonography, in

K-8 conjunction with serum analysis, has become a powerful
tool in the assessment of risk for aneuploidy, in both the
Different clinical presentations of endometriosis
first and the second trimester. In the mid trimester the

Asefjah H. diverse sonographic patterns seen in the different
Parsian Hospital, Tehran, Iran. aneuploidies allows clinicians to guide patients to a
Email: [email protected] presumptive diagnosis. The information obtained
noninvasively helps the expectant couple to weigh the
Endometriosis is the third leading cause of gynecologic risks of invasive testing against the probability of
hospitalization in the United States. Endometriosis can having a child with an abnormality. The goal of
develop between 10 and 60 years of age. The average screening is the detection of a greater number of
age of diagnosis is 27 years. This disease impacts both a karyotypically abnormal fetuses with fewer invasive
woman‟s physical and mental wellbeing. This impact is procedures and subsequently the loss of fewer normal
often compounded by the frequent delay of 6 years or fetuses. First and second trimester markers will be
more from the onset of symptoms to a confirmed discussed.
diagnosis, which may. Because there is no good
noninvasive test for endometriosis, there is often a K-10
significant delay in diagnosis of this disease. Among
Religion, law and ethics: Ethical and
women who seek tubal ligation, the prevalence of
anthropological reflection on assisted
endometriosis appears to range from 2-18%, whereas
reproduction
within infertile populations it has been reported to be as

high as 50%.
Garmaroudi Sh.
No serum marker has been found to diagnose University of Zurich, Brocher Foundation, Zurich, Germany.
endometriosis with adequate sensitivity and specificity. Email: [email protected]
There has been a recent focus on the presence of nerve
fibers in the eutopic endometrium of patients with This paper seeks to have a close look at what happens
endometriosis. There is a wide spectrum of symptom with cosmological phenomena, economic values,
severity, clinical presentation and the stage of conflicting moralities and kinship principles when they
endometriosis. Laparoscopy is the gold standard for meet clinical practices, legislations and regulations
diagnosis of endometriosis but in the hands of expert pertaining to reproductive technologies, including
laparoscopic surgeon 6-10% of endometriosis is missed. gamete and embryo donation as well as surrogacy
Some patients with minimal disease have debilitating arrangements. Mainly, based on my extensive
pain, whereas other women with severe stage III to IV ethnographic research on assisted reproductive
disease are asymptomatic. Women with mild to technologies in Iran, which includes an examination of
moderate endometriosis have a higher incidence of the normative arguments, this paper attempts to explore
endocrine abnormalities, anovulation, corpusluteum what moral, theological and legal reasoning underlie the
insufficiency, hyperprolactinemia, luteinized unruptured concepts of kinship and reproduction that move people
follicle syndrome, and spontaneous abortions. when they turn to- or refrain from- certain technologies
4 Iranian Journal of Reproductive Medicine, Vol. 13, No 4 (Suppl. 1), April 2015
Abstracts of the 6th Yazd International Congress and Student Award in Reproductive Medicine with 1st Reproductive Genetic Congress
of assisted conception? And how are they contested and more study on this subject is necessary to elucidate the
negotiated? I acknowledge the importance of contextual best compound and the optimal doses and duration of
understanding of moral concepts, arguments and these medications.
reasoning involved in the application of reproductive
technologies as well as the interplay between religious, K-12
moral and legal ideas and institutions, and the place of The sperm aging: Is it affecting ART outcomes?
this interplay in contemporary debates surrounding
human reproduction and reproductive health. I view Sadeghi MR.
reproduction as a process through which the Department of Embryology and Andrology, Reproductive
Biotechnology Research Centre, Avicenna Research Institute,
foundational structures and perceptions of a society and
Tehran, Iran.
its dynamics are reproduced and contested rather than a
Email: [email protected]
sexual act or as simply the combination of male and
female reproductive substances. Reflections offered in Sperm aging is usually the concerns of many basic and
this article are based on my doctoral research project clinical studies recently. This topic can be considered
“Assisted Reproductive Technologies in Iran from an from three different perspectives. Most of studies are
Anthropological Perspective: Legal and Jurisprudential focused on the paternal age on quality and quantity of
Responses and Social Dynamics” that has examined the sperm and also its consequences on the older men
Iranian and contemporary Shia legal debates and fertility, in vitro fertilization outcomes and the health
discussions on technologies of assisted conception and status of infants born from these parents. Its second
has looked at the regulations and implementation of aspect is sperm aging in time interval between
these technologies in Iran. spermiation and ejaculation. Based on the tracing of
radiolabeled molecules, usual duration of sperm journey
K-11 is approximately two weeks, but it is highly affected by
Antioxidants and their role in male infertility time interval between ejaculations. The third aspect of
sperm aging is lapse of time from ejaculation. Human
Moein MR. sperm exposed to a physicochemical condition
Research and Clinical Center for Infertility, Shahid Sadoughi
following ejaculation in a container that are very
University of Medical Sciences, Yazd, Iran.
different from in vivo condition of male and female
Email: [email protected]
genital tracts. These physicochemical changes can lead
Male infertility constitutes about 50% of cases of to deleterious consequences on sperm structure and
infertility. In nearly half of these patients no specific function. Several studies on aged men in comparison
etiological cause could be found. In recent decades younger ones showed that the spermogram parameters
much attention has been paid to the role of decreased significantly in older men. Sperm chromatin
overproduction of free radicals in semen and oxidative integrity and DNA fragmentations, as well as
stress on different sperm functions. Also sperm aneuploidy abnormalities significantly increased with
chromatin damage which could be a result of oxidative men‟s age.
stress or other known factors such as varicocele or In infertile men with oligoasthenoteratozoospermia
smoking has been implicated in etiology of male (OAT) too long intervals between ejaculations lead to
infertility. With respect to these new findings removal decrease sperm quality. However multiple ejaculations
of free radicals and elimination of factors that could in short interval in these patients significantly increased
potentially induce sperm chromatin damage became the sperm parameters and its chromatin integrity. There
more important. With regard to the above mentioned is negative correlation between the ejaculation to
findings, during the past few years a variety of different analysis, processing and insemination intervals. So that
antioxidant medications and even foods rich in sperm parameters significantly declined during in vitro
antioxidant compounds had been tried to treat these storage of sperm especially following processing and
patients. Many studies and reports evaluated the effect elimination of seminal plasma.Therefore, ignoring of
of these compounds on improving sperm parameters the sperm aging in relation to human fertility, especially
and improving pregnancy rate. Some of these studies assisted reproductive technologies can have serious
showed the positive effect of antioxidants on fertility influences on natural fertility, IVF outcomes and the
potentials while other observed no significant effects. health of associated offspring.
Also there is a still controversy on the best type of
antioxidant and even the optimal dose of these K-13
compounds in the treatment of male infertility, which New markers in male fertility evaluations
needs more clinical trials and further studies. In
conclusion it seems that antioxidants have an important Talebi AR.
Research and Clinical Center for Infertility, Shahid Sadoughi
role on improving male infertility and its use in patients
University of Medical Sciences, Yazd, Iran.
with idiopathic male infertility is strangle advised, yet
Email: [email protected]; [email protected]
Iranian Journal of Reproductive Medicine, Vol. 13, No 4 (Suppl. 1), April 2015 5
Abstracts of the 6th Yazd International Congress and Student Award in Reproductive Medicine with 1st Reproductive Genetic Congress
It is obvious that the assessments of spermatogenesis, 2. John van Geest Cancer Research Center, Nottingham
semen and endocrine analysis are critical in the Trent University, Nottingham, UK.
evaluation of the infertile and subfertile couples. But, it Email: [email protected]

seems that currently used methods are inadequate to
Introduction: In many female species, sperm viability
correctly predict sperm fertility potential and do not
is maintained by storing spermatozoa in the
provide sufficient information for diagnosing and
reproductive tract prior to fertilization via the temporary
treatment of some clinical infertility situations.
attachment of sperm heads to the apical oviduct
Conversely, the hidden and unclear sperm abnormalities
epithelial cells (OECs). Despite its importance in
impairing the reproductive success of sperm and egg
assisted reproduction, the mechanisms involved in the
interaction often remain undiagnosed and in these cases
prolongation of sperm survival in vivo are not fully
of unexplained infertility, there are no clear reasons for
understood. It has been reported that the presence of
the condition. So, many laboratory tests have been
sperm in the female oviduct induces the expression of
developed in order to evaluate the structure and function
the constitutive member of the 70 kDa heat shock
of human spermatozoa.
protein family, HSPA8, in the oviduct as an exogenous
In recent years, researches have focused on identifying
protein and we have shown that exogenous recombinant
reliable markers of fertility at the genomic, proteomic,
HSPA8 enhances survival and membrane fluidity of
biochemical, and immunocytochemical levels. The use
boar spermatozoa in vitro. The aim of this study is to
of fluorescent markers to assess the acrosomal status,
provide insight into the capacity of exogenous HSPA8
the use of vital staining for mitochondrial activity and to extend sperm survival and mechanism of HSPA8-
energy metabolism and the use of particular sperm interactions.
fluorochromes and cytochemical dyes to detect altered Materials and Methods: The localization of
sperm chromatin or DNA along existing functional tests fluorescently conjugated exogenous HSPA8 (ATTO488-
like the hypo-osmotic swelling test and the hemi-zona HSPA8) following incubation with boar spermatozoa
assay are the most useful assessments of spermatozoa. (0.5 µg/ml, 15 min, room temperature) was determined
Additionally, an association between infertility and by confocal microscopy. Sperm viability (membrane
seminal oxidative stress has been suggested. Excessive integrity) was assessed using SYBR-14/propidium
ROS production damages the sperm membrane, reduces iodide. Membrane fluidity (D values and Recovery %)
motility, induces permanent DNA damage and it is of acrosomal and posacrosomal domains of live cells
closely associated with apoptosis. ROS production can was measured using fluorescence recovery after
be directly monitored by a luminol or a lucigenin-based photobleaching (FRAP). The influence of membrane
chemilluminescence assay and the apoptosis can be cholesterol on the ability of HSPA8 to modulate sperm
detected by several molecular and immunocytochemical membrane fluidity was examined by depleting
methods. In many cases of male infertility, the cause is membrane cholesterol using different concentrations of
genetically in origin. cyclodextrin (0, 2, 4, 8 mM, 30 min) and replenishing
Thus, in the context of reproductive research, genetic cholesterol using cyclodextrin-cholesterol complexes.
defects in gametogenesis are being extensively studied Data are expressed as mean±SEM.
and many important genes in sperm biology have Results: ATTO488-HSPA8 binding was localized to the
detected so far. Finally, the proteomics or acrosomal sperm membrane and HSPA8 had no effect
comprehensive study of proteins with their particular on the viability of cholesterol-depleted spermatozoa.
structural and functional aspects, have allowed the Cholesterol removal reduced D values for the acrosome
identification of different proteins in semen and (46±3 vs. 29±2, p<0.01) and postacrosome (34±6 vs.
spermatozoa that are responsible for the regulation of 19±4, p<0.005). R% values were also significantly
normal/defective sperm functions. Presently, numerous lower. Reloading cholesterol restored membrane fluidity
proteomics techniques, such as two-dimensional (2D) and the ability of HSPA8 to increase viability.
polyacrylamide gel electrophoresis, and mass Conclusion: Spermatozoa are devoid of protein
spectrometry are widely used to identify sperm-specific synthesis apparatus and incapable of de novo protein
proteins. These assays help us to understand different synthesis under stressful conditions. These findings
functional aspects of sperm proteins which are suggest that exogenous heat shock proteins can maintain
important in motility, capacitation, acrosomal reaction, the integrity of biologic membranes and act as „rapid
response‟ extracellular cytoprotectors in a cholesterol-
fertilization, chromatin remodeling and posttranslational
modifications. dependent manner.

K-15
K-14
Pluripotent stem cells and regenerative
Exogenous HSPA8 prolongs sperm survival by
medicine
enhancing membrane fluidity in a cholesterol-

dependent manner
Veiga A.

Banc de Línies Cel·lulars, Centre de Medicina Regenerativa
Moein Vaziri N1, Holt WV1, Pockley AG2, Fazeli AR1. de Barcelona, Servei de Medicina de la Reproduccio, Hospital
1. Academic Unit of Reproductive and Developmental Universitari Quiron Dexeus, Barcelona, Spain.
Medicine, The University of Sheffield, Sheffield, UK. Email: [email protected]
6 Iranian Journal of Reproductive Medicine, Vol. 13, No 4 (Suppl. 1), April 2015
Abstracts of the 6th Yazd International Congress and Student Award in Reproductive Medicine with 1st Reproductive Genetic Congress
Introduction: Pluripotent stem cells have been defined include the most frequent haplotypes have to be put in
as those with the ability to give rise to all tissue types place. iPS generation for specific patients or from cell
from the three embryonic germ layers (ectoderm, types previously banked, such as cord blood cells, may
mesoderm, and endoderm) and the capacity for solve the problem in reprogrammed cells. A clinical
indefinite self-renewal if cultured in appropriate trial involving the use of hESC derived Retinal
conditions. Two different cell types, embryonic stem Pigmented Epithelium for Macular degeneration is
cells (hESC) and induced pluripotent stem (iPS) cells; currently in place in the US and UK and the same
have been demonstrated to be pluripotent. protocol have been approved with iPS cells in Japan.
Human embryonic stem cells: Human embryonic stem The field of stem cell research and regenerative
cells (hESC) were first described in 1998 by Thomson medicine holds a promising future for the treatment of
and are derived from human embryos, mainly from the degenerative diseases.
Inner Cell Mass (ICM) of the blastocyst, at day 5-7 of
development. Other options for hESC derivation include K-16
early embryos, morulae and single cells. Embryos Next steps towards the transplantable artificial
donated for research by couples undergoing In Vitro ovary
Fertilization treatment constitute the main source for
hESC derivation. The methodology may vary among the Amorim CA.
different groups and no standardized protocol for Pôle de Recherche en Gynécologie, Université catholique de
Louvain, Brussels, Belgium.
derivation has been described. To date, more than one
Email: [email protected]
thousand hESC lines have been derived worldwide and

even though an international registry is still lacking,
In recent years, advanced chemo/ radio therapeutic
more than 600 European and international hESC lines
treatments have led to high survival rates in cancer
have been registered at the human Embryonic Stem Cell
patients, giving rise to new issues for cancer survivors.
registry, a project funded by the FP6 work programmer
Indeed, one major concern is future fertility in these
of the European Commission.
women, since they may face premature ovarian failure.
Induced pluripotent stem cells (iPS): In 2006,
For this reason, different strategies have been proposed
Yamanaka and co-workers described the possibility of
to preserve their fertility. When gonadotoxic treatment
reprogramming the nucleus of mouse somatic cells into
cannot be delayed, ovarian tissue cryobanking appears
a pluripotent state by the ectopic expression of a defined
to be the most promising way of preserving a patient‟s
set of genes. These cells were called induced pluripotent
fertility. Moreover, this is the sole means of
stem cells (iPS). A year later, 2 different reports
safeguarding fertility in prepubertal girls. Auto
described the methodology to generate human iPS by
transplantation is the only option able to reestablish
retroviral transduction of 4 different sets of genes (Oct ,
4 ovarian function from cryopreserved ovarian tissue in
Sox , Klf and c-Myc and Oct , Sox , Nanog and
2 4 4 2 cancer survivors at present. So far, this technique has
Lin28). These cells exhibit most of the characteristics
led to successful ovarian function restoration and up to
seen in hESC, such as morphology, proliferation ability
40 pregnancies in a number of centers around the world.
and pluripotency. A number of publications have
However, there is a legitimate concern regarding the
demonstrated that somatic cells from different origins
possible presence of malignant cells in frozen-thawed
can be reprogrammed to iPS (fibroblasts, keratinocytes,
fragments, which could provoke a recurrence of the
liver cells, neural stem cells, cord blood cells, etc.) with
primary disease after re-implantation. Although many
the use of a limited number of transcription factors.
types of cancer never metastasise to the ovaries,
Also, the mode of delivery of such factors has been
leukaemia is systemic in nature and poses a greater
modified to achieve safe reprogramming.
threat to the patient, while breast cancer and some types
Clinical translation of pluripotent stem cells: There
of lymphoma are classed as moderate risk.
are a number of major drawbacks that need to be
For these patients, a safer alternative could be grafting
resolved to ensure the safe application for therapy of
of isolated preantral follicles, as these structures are
pluripotent stem cells, including hESC and iPS. One of
enclosed in a basement membrane that prevents direct
the major issues to be solved is to determine which cells
contact between follicular cells and capillaries, white
have to be transplanted, and specifically at what stage of
blood cells, and nerve processes. Since ovarian cells
differentiation. Also, when transplanting into solid
(OCs) are essential for follicle development and
organs, the 3D support for transplantation and
neovascularization, autologous OCs should be grafted
integration also has to be considered. Differentiation
together with isolated follicles. To replace the original
protocols have to be optimized in order to produce pure
ovarian structure, a transplantable artificial ovary
populations. Large scale and GMP production of cells
(TAO) should be created in order to encapsulate and
are required. Similar to organ transplantation, immune
protect the isolated follicles and OCs. As in case of a
rejection should also to be considered. hESC banks that
natural ovary, the main goal of the TAO is to offer an
Iranian Journal of Reproductive Medicine, Vol. 13, No 4 (Suppl. 1), April 2015 7
Abstracts of the 6th Yazd International Congress and Student Award in Reproductive Medicine with 1st Reproductive Genetic Congress
environment that allows follicle survival and In this study we evaluated presence and amount of: a)
development. Therefore, a TAO should maintain the ooplasmic vacuolization, b) organelle-specific
original structure of follicles, ensure proper associations such as mitochondria-smooth endoplasmic
communication between follicles and OCs, and preserve reticulum (M-SER) aggregates and mitochondria-
their interaction with the extracellular matrix and supply vesicle (MV) complexes, and c) cortical granules (CGs).
factors involved in follicular survival and development. Materials and Methods: MII oocytes were subjected to
In other words, the TAO should spatially and slow freezing through two-step propanediol (PrOH)
temporally mimic the ECM. In order to do so, it should dehydration with 0.75-1.5 mol/l PrOH and 0.2 mol/l
include some design parameters, such as physical sucrose and examined by light and transmission electron
support of follicles, porosity, bioactivity, microscopy (TEM) at different time intervals during
vascularization, interaction with cells, and thawing. Cryopreserved oocytes were fixed after being
biodegradability, which are all interconnected and transferred in 1.0 mol/l PrOH and 0.3 mol/l sucrose
influence each other. (group A, n=15), 0.5 mol/l PrOH and 0.3 mol/l sucrose
(group B, n=15) and 0.3 mol/l sucrose (group C, n=15).
K-17 Fresh MII oocytes (n=15) were used as controls.
Mitochondria and oocyte maturation Results: Morphometric and TEM analysis revealed that
vacuoles were only occasionally detected in the
Salehnia M. ooplasm of fresh controls. Conversely, vacuoles were
Department of Anatomy, Tarbiat Modares University, Tehran,
numerous in the cryopreserved oocytes of group A and
Iran.
appeared to reach an even larger number in group B
Email: [email protected]; [email protected]
oocytes. M-SER aggregates, large and abundant in the
Mitochondria are critical organelles within the cell and ooplasm of fresh controls, significantly decreased in
has important role in the oocyte development. The number following freezing, particularly in the oocytes
number and distribution of mitochondria, and energy belonging to groups A and B. MV complexes were
(ATP) production are critical factors that influence not instead small and scarce in fresh control oocytes but
only on the maturation and development of the oocyte augmented after freezing, being especially abundant in
but also on its fertilization, and subsequent embryo the oocytes belonging to group B. Vacuoles and MV
development. Structural and metabolic mitochondrial complexes both diminished in the oocytes belonging to
defects are associated with failures in oocyte maturation group C, whereas M-SER aggregates increased in
and abnormal development or arrest of embryos. number. CGs was scarce in all cryopreserved oocytes in
Mitochondrial content could affect the fertilization respect to those found in fresh controls and gradually
potential of oocyte. If mitochondrial DNA content of diminished as thawing progressed.
oocyte be lower than threshold, it‟s more prone to failed Conclusion: This study proves that vacuoles, generally
maturation and showed reduced fertilization rates. regarded as markers of oocyte cryodamage during slow
Dysfunction of oocyte mitochondria may occur without cooling, may form during freezing, but become
detectable morphological abnormalities. numerous during thawing, particularly when the lowest
concentration of PrOH is reached. Significant variations
K-18 in the number of M-SER aggregates and MV complexes
Organelle morphodynamics in human mature occurred during the freeze-thawing, suggesting a
oocytes after cryopreservation. Ultra structural dynamic process of transition between these two forms
of organelle associations. This study also evidences that
analysis at different time intervals during
a premature CG exocytosis progressively occurs during
thawing
the whole freeze-thawing procedure. It seems also worth

Nottola SA1, Borini A2, Levi-Setti PE3, Macchiarelli G4. noting that all systems of ooplasmic membranes appear
1. Department of Anatomy, Histology, Forensic Medicine significantly concerned by freeze-thawing but, except
and Orthopaedics, Sapienza University, Rome, Italy. for CGs, their alterations seem to undergo a partial or,
2. Tecnobios Procreazione, Center for Reproductive Health,
more rarely, an almost complete recovery at the end of
Bologna, Italy.
the thawing process.
3. Department of Gynecology and Reproductive Medicine,

IRCCS Istituto Clinico Humanitas, Rozzano, Milan, Italy.
4. Department of Life, Health and Environmental Sciences, K-19
University of L´Aquila, L’Aquila, Rome, Italy. Ultrastructural markers of aging in human
Email: [email protected]
oocytes

Introduction: During freeze-thawing, the human
Macchiarelli G1, Bianchi S1, Micara G2, Aragona C2,
metaphase II (MII) oocyte is exposed to a variety of
Nottola SA3.
physical and chemical conditions that may endanger its
1. Department of Life, Health and Environmental Sciences,
competence to fertilization and even its mere survival. University of L´Aquila, Rome, Italy.
8 Iranian Journal of Reproductive Medicine, Vol. 13, No 4 (Suppl. 1), April 2015
Abstracts of the 6th Yazd International Congress and Student Award in Reproductive Medicine with 1st Reproductive Genetic Congress
2. Department of Gynecology-Obstetrics and Urology, cells are removed, but removing more than one cell can
Sapienza University of Rome, Rome, Italy. cause more damage to the embryo. There are much
3. Department of Anatomy, Histology, Forensic Medicine
debates about the number of cells, the embryonic stage
and Orthopaedics, University of Rome, Rome, Italy.
Email: [email protected] and the technique of biopsy. Quality control after biopsy
is most important issues in this regard. Naturally before
Introduction: The delay of childbearing contributes to implantation, embryo hatched from zona pellucid,
the increasing proportion of subfertile couples which in some cases, such as aging and freezing, this
necessitating assisted reproduction technology (ART) hatching does not happen. So assisted hatching can be
procedures. Subfertility relates with decay in oocyte helpful in these cases. Removal of degenerated cells
quality due to reproductive aging, indeed maternal aging from fresh embryos or embryos after thawing in some
impairs reproductive potential. Prolonged culture, also cases, may be helpful to keep a better growth of the
called “in vitro aging” may also impair oocyte embryo. Transferring the nucleus, cytoplasm and
competence. Ultra structural oocyte quality greatly mitochondria of healthy oocyte in the oocyte case is
affects ART outcome that also depends on to specific currently being done in some countries. It should be
morphological parameters. In this report, we account for noted that many other manipulations such as human
the ultrastructural markers of aging, in oocytes from cloning need the legal and ethical permission.
over-35 years old women underwent to ART
procedures, enrolled in this study after informed K-21
consent. Foreseeing the fate of the embryo- Advances
Materials and Methods: We studied MII oocytes from and limitations of time-lapse technology
women under 35 and over 35 years old, fixed at pick up
or after 24 hr culture. Ultrastructural and morphometric Losonczi E.
Department of Genetics, Faculty of Veterinary Sciences, St.
evaluations were performed.
Istvan University, Hungary.
Results: Significant increasing of vacuoles, decreasing
Email: [email protected]
of mitochondria-smooth endoplasmic reticulum
aggregates, increasing of mitochondria-vesicle Embryo evaluation is a crucial part of the infertility
complexes density, decreasing of cortical granules and treatment. It supports the selection of the right embryo
microvilli, increasing of zona pellucida density and from the cohort for transfer if performed properly. A
thickness, characterized oocytes from aged women. good scheme for embryo assessment assists elective
These changes were more evident in the oocytes single embryo transfer, regardless if a clinic utilizes
submitted to prolonged culture. fresh and cryopreserved transfers or follows the “freeze
Conclusion: These changes may be assumed as ultra- all” strategy. Moreover, a proper evaluation system
structural markers of oocyte aging. It was also coupled with proper embryo culture conditions and
demonstrated that oocytes from younger women are less manipulation techniques can have an effect on
sensitive to prolonged culture (in vitro aging) than the stimulation protocols in favor of mild approaches,
oocytes from aged women. resulting in lower hormonal load, fewer but higher
quality embryos, and, consequently, higher embryo
K-20
utilization.
Manipulation of human oocytes and embryos to Embryo evaluation techniques have been developed
diagnose and treat from point of embryologist from the advent of mammalian embryology. These were
view based on static evaluation of the morphology at certain
time-points during in vitro culture. Scores were
Ghaffari Novin M. established to evaluate certain morphological features
Department of Anatomical Sciences and Biology, Faculty of
including pronuclear pattern, zona pellucida, blastomere
Medical Sciences, Shahid Beheshti University of Medical
number at certain time-points, extent of fragmentation,
Sciences, Tehran, Iran.
Email: [email protected] cytoplasmic appearance, and blastocyst morphology.
For this, embryos had to be removed from the incubator
With the birth of the first human baby by IVF and checked. How often? Preferably not at all, but to get
techniques in 1978 AD, this area of medicine is in very any information about embryo quality during in vitro
rapid advances in the diagnosis and treatment of development one needs to make compromises, so
infertility. An experienced clinical embryologist using frequency for embryo checkups generally range from 1-
advanced facilities can perform various manipulations 5 times during the 5 days of culture.
on oocytes and embryos. Chromosomal and genetic One of the advances of using time-lapse techniques is to
analysis of oocytes and embryos biopsied cells could be follow up embryo development and to obtain
helpful in the diagnosis of many diseases. To avoid false information on morphology. This information is
reports because of mosaicism, geneticists suggest two provided continuously every 5-20 min, not just at
Iranian Journal of Reproductive Medicine, Vol. 13, No 4 (Suppl. 1), April 2015 9
Abstracts of the 6th Yazd International Congress and Student Award in Reproductive Medicine with 1st Reproductive Genetic Congress
distinct time-points of the day but at any time-points of Fragmentation has also been observed as highly
the day. Techniques now available make it possible to dynamic process with fragments being continuously
perform embryo evaluation while embryos are inside of rearranging around the blastomeres or being reabsorbed
the incubator, thus reducing handling stress. During a during the course of in vitro development. For this
course of routine time-lapse examination, hundreds of reason, static evaluation of fragmentation might not be
images are made and saved, archived in digital format, absolutely correct. A further example for the dynamic
enabling another fundamentally important expectation nature of morphology is the fact that blastocysts pulsate:
of the scientific society: proper documentation and they expand and collapse continuously. An expanded
quality control of the laboratory phase, right at its heart: blastocyst may collapse within a short time, whereas her
inside of the incubator. Apart from quality control there blastocyst score would change, while her quality would
is another everyday use of the digital imaging, and that not.
is learning, teaching and communication. The listed Time-lapse projects also provide insight into the timings
possibilities alone justify the use of time-lapse of the cell cycle. Embryos are supposed to cleave within
technology in the embryology lab. However, the a definite time-frame. Which are the most important
exponentially increased number of information provided events, and are they in correlation with blastocyst
by time-lapse technology has put a question mark onto formation of pregnancy? Recent studies have revealed
the reliability of the well-established morphological that cleavages up until the 4 cell stage are more
scorings. Moreover, a new set of information became predictive to the chance to reach the blastocyst stage,
available, as time-lapse enables us to calculate with the while events prone to happen after the onset of the
length of interphases and the duration and synchrony of genomic activation seem to provide information that is
cytokinesis, and use this information when quantifying relevant to pregnancy. According to our group,
embryo quality. How does time-lapse technology morphokinetics in itself is not sufficient for proper
change how see and grade embryos? embryo evaluation; it has to be applied in combination
What do we learn from continuous embryo follow- with static morphology. Nevertheless, time-lapse is
up? needed to qualify static morphology properly. Focusing
Pronuclear scoring involves the assessment of the onto the importance of morphokinetics purely, our
number and relative position of the nucleolar precursor group sees its role in supporting de-selection. De-
bodies (NPBs) which are established in the pronuclei. selection in this content means embryos performing
Any inequality in the distribution of the NPBs within irregular cleavage like directly cleaving from one cell to
the pronuclei is considered to be abnormal, but time- three cell stages are ranked back in the cohort with the
lapse studies reviled, that NPBs move around inside of note of lower chance for implantation.
the pronuclei, and can produce up to 2 score difference Up to date there are numerous equipment available that
within 2 hr, making their traditional, “static evaluation” can host live cells and follow up their development
and its value questionable in the present format. while maintaining and supporting close to physiological
Morphologically, early cleaving embryos have been environment around the cells. The most classical type of
regarded as higher quality ones. However, early equipment designed for live cell imaging is a regular
cleavage has lost its classic meaning in the time-lapse inverted microscope with a plastic cask built around it.
environment. At checking time-lapse recordings of In the plastic box temperature, humidity and gas
embryos we are looking for timeframes for cleavages, concentration can be adjusted up to certain precisity,
as too early cleavage can equally be an unfavorable sign with uneven distribution. An alternative solution is to
of embryo quality as cleaving too late. Such simple apply a small incubation box (stage-top incubator) onto
questions as cell number at certain time-points get also the microscopic stage. This type of equipment, available
questioned, when it became possible to follow the from all major microscope manufacturers and also
cleavage pattern of the actual embryo. A home-made editions do not satisfy the delicate needs of
morphologically sound five cell stage embryo can reach the embryo for a precise and stable environment.
the five cell stage by normal but also abnormal cleavage However, with the use of an environmental chamber in
paths. A five cell stage embryo can be the result if the combination with a stage-top incubator made it possible
first cytokinesis produced 3 blastomeres, 2 of which to follow up human embryo development until the 4 cell
cleaved further. After a normal first cytokinesis, one stage. Further developments included the inclusion of a
blastomere can cleave to 2 cells, while the second one proper, automated inverted microscope into a regular
may cleave to 3 daughter cells, resulting, again, in a incubator. This setup needs a quasi-robotic system that
normal looking but abnormal five cell stage embryo. moves the embryos into the field of view. Besides the
Though the morphological evaluation may reveal same adequate optical output, the efficacy of the earlier
score for the given examples, their potential to implant versions were hampered by the complicated inner
and to develop to a healthy offspring differ structure conveying heat accumulation due to friction,
significantly. VOC due to lubrication, sheer stress due to movement,
10 Iranian Journal of Reproductive Medicine, Vol. 13, No 4 (Suppl. 1), April 2015