Assisted Reproductive

Technology (ART) and

 Application 

Yu-Hui Tsai, Ph.D.

Graduate Institute of Medical Sciences

Taipei Medical University
 Infertility
• Couple who does not use any
contraceptive, but can not conceive
within 2~3 years are considered
infertile.
• Up to 5,000,000 American couples suffer
from infertility every year and 20% of
those decide to undergo in vitro
fertilization.
• About one out of every 6 couples is
infertile in the US, and Taiwan as well.
 Male infertility
• Infertility due to male factor accounts for at
least 40% of infertility cases. For this reason,
it is very important that men also be
investigated for fertility problems. While
blockages and problems producing healthy
sperm are often the main reasons for male
infertility, there are a variety of issues that
can diagnosed.
• Once a diagnosis has been made by a
fertility specialist, a proper course of
treatment can be recommended. Depending
on the cause of male infertility, possible
forms of treatment may include surgery,
drugs, or assisted reproductive technologies,
like ICSI.
 Decreasing male fertility rates

• Recent studies have identified several factors

causing a decline in male fertility:
• The anti-impotence drug Viagra could be
damaging sperm acrosomes, and lowering the
taker’s ability to conceive (40% less). (Fertility
and Sterility).
• Another study suggests that pollution from
chemicals such as dioxin can lower a man's
sperm count. (in the environmental journal the
Ends Report)
• Endocrine-disrupting chemicals are also
thought to be affecting fertility rates.
Menevit, the 'first ever drug for male
infertility',

• Menevit, developed in Australia, contains

antioxidants which works by acting on free
radicals that fragment sperm, the main
cause of infertility.
• In a preliminary study of 60 infertile men,
the rate of pregnancy was increased
significantly, but larger clinical trials are
required before the drug can be merited.
 Female infertility
• Ultrasound is likely to be done first, as this can
alert your specialist to any possible problems.
There are also a number of blood tests that can
be performed. In women, the most common
ones include:
• Day 3 FSH testing
• Luteinizing hormone (LH) testing
• Progesterone testing
• Estradiol Level testing
• Hysterosalpingogram
• For women who are suspected of having low
ovarian reserves, a Day 3 Inhibin and a
Clomiphene Challenge test may be performed.
• Antibody against husband’s sperm
 Treating Infertility: Assisted
Reproductive Technology (ART)
• A large percentage of couples face difficulties in
getting pregnant, many have found success with
Assisted Reproductive Technology (ART).

• ART involves a number of different procedures

to help address fertility problems and increase
the likelihood of pregnancy. Be sure to speak
with the reproductive endocrinologist about ART
procedures that may be right for you to increase
your chances of conceiving.
 Assisted Reproductive
Technology (ART)

• Insemination
sperm collection/ sperm donation
• In vitro fertilization
egg retrieval/ egg donation
• Surrogate
 Who needs IVF?

• Couple has infertility problem.

– Men with low sperm count, abnormal sperm, no
mature sperm, antibody against sperms etc.
– Women with oviduct obstruction, endometriosis,
endocrine imbalance, reproductive organ infection,
antibody against partner’s sperms etc.

• The couple both carry recessive genetic

disorder on the same genetic locus, or
female with X-linked genetic disorder .
In vitro fertilization (IVF)
( 試管嬰兒 )
 In vitro fertilization (IVF)
( 試管嬰兒 )

• IVF is the uniting of egg and sperm in vitro

(in the lab). Subsequently the embryos are
transferred into the uterus through the
cervix and pregnancy is allowed to begin.
 In vitro fertilization (IVF)
( 試管嬰兒 )
• IVF is a major treatment in infertility when other
methods of assisted reproductive technology have
failed. The process involves
– ovulation induction through hormone treatment,
– monitoring of hormone levels and follicle scans with
ultrasound,
– egg retrieval from the woman's ovaries
– and fertilize eggs with sperms in a fluid medium.

• The fertilized egg (zygote) is then transferred to

the patient's uterus with the intent to establish a
successful pregnancy.
 Procedures for IVF
• Day 0 Egg retrieval, Sperm collection, and
preparation
• Insemination in vitro
• Day 1 Check eggs for fertilization (the presence
of two pronuclei or PN's)
• Day 2 Embryos at the 4-cell or more stage of
development
• Day 3 Embryos at the 8-cell or more stage of
development
• Day 4 Embryos at the compacted morula (16-32
cell) stage
• Day 5 Embryos at the blastocyst stage of
development
 Inducing Superovulation
• Injections of Lupron, Metrodin, Pergonal or
Humegon allow the ovaries to develop many
follicles as seen in this ultrasound immediately
prior to egg retrieval.
Retrieved egg
Human MI oocyte with extruded polar body
Human MII oocyte
Human MII oocyte with prominent spindles
 Fertilization of retrieved egg
• Retrival of maturing eggs via laparoscopy.
• Each egg is incubated with > 3.5 x 106 sperms.
The fertilized egg contains two pronuclei. Each
contains half the chromosomal material from
each one of the genetic parents.
 Embryo at 4-cell stage
• A four cell pre-embryo. Each of the cells is
called a blastomere. The embryo is
surrounded by a protein matrix "shell"
called the zona pellucida.
 Transferring of 8 cell-stage embryo
• On the third day after egg retrieval eight
cell pre-embryos can be transferred to the
uterus. On average, 15 to 25% of embryos
will implant after being transferred.
Hatching in vivo prior to implantation
• Prior to implantation, the pre-embryo, now at the
blastocyst stage, must hatch out of the zona
pellucida prior to implantation.
Intracytoplasmic sperm
injection (ICSI)
 Intracytoplasmic sperm
injection
• Intracytoplasmic sperm injection (ICSI,
pronounced "eeksee") is an in vitro fertilization
procedure in which a single sperm is injected
directly into an egg.
• This procedure is used to overcome male
infertility problems, although it may also be
used where eggs cannot easily be penetrated by
sperm, and occasionally as a method of in vitro
fertilization, especially that associated with
sperm donation.
 How is ICSI performed?
• Using micromanipulation technology, ICSI
allows fertility specialists to fertilize an egg
using just one sperm. While it is preferred to use
sperm from a semen sample, specialists can
retrieve sperm from the testicles if it is necessary.
Once sperm has been collected, the specialist will
draw a single sperm into a needle and inject it
directly into an egg that has been collected from the
female partner through the usual retrieval methods.
• This process bypasses the conventional IVF
methods of fertilization, thereby ensuring that
fertilization has taken place. The fertilized eggs
are then left to culture for a few days before being
transferred back to the woman’s uterus. ICSI is
always used alongside IVF.
 Intracellular
sperm injection
(ICSI)
 Who needs ICSI?
• Men who have very poor semen quality or
azoospermia should be offered ICSI.
– Low sperm count
– Poor sperm motility
– Abnormally high amount of morphologically
atypical sperm
– Experienced fertilization failure for unknown
reasons in a previous IVF cycle
– Lack of any sperm in ejaculate due to CAVD,
failed vasectomy reversal, failure to produce
sperm, or an obstruction in the epididymus due to
past inflammation
– Retrograde ejaculation
– Immunological factors
Gamate/embryo transfer
• Many of the large programs have attempted
blastocyst culture, but returned to day 3
transfers because they had trouble getting
the embryos to grow the blastocyst stage.
Growing the embryos to the blastocyst
stage requires great attention to detail - a
luxury not afforded in a big program
performing dozens of procedures a day.
• Programs performing less than 200 cycles
per year have had the best luck with
blastocyst culture and transfer with
reported (but unverified) pregnancy rates in
the 50-70% range for younger patients and
egg donor cycles.
Gamete intrafallopian transfer
• In gamete intrafallopian transfer (GIFT),
eggs are removed from the woman, and
placed in one of the fallopian tubes,
along with the man's sperm. This
allows fertilization to take place inside
the woman's body. Therefore, this
variation is actually an in vivo
fertilization, and not an in vitro
fertilization.
Zygote intrafallopian transfer
• Zygote intrafallopian transfer (ZIFT) is an
infertility treatment where a blockage in
the fallopian tubes are the cause. Egg
cells are removed from a woman's ovaries,
and in vitro fertilized. The resulting zygote
is placed into the fallopian tube by the use
of laparoscopy.
IVD: ivfed eggs developed in vivo
• A device that allows IVF embryos to develop in
the womb rather than a laboratory dish,
developed by Swiss company Anecova, is to be
trialed at CARE Fertility in Nottingham.
• The treatment, named in vivo development, or
IVD, has been termed a more natural alternative
to IVF and there are hopes that it will lead to
fewer embryos with genetic abnormalities being
produced.
• In normal IVF eggs are fertilized with sperm and
allowed to grow for a few days in a laboratory
dish containing chemicals and nutrients, before
being transfered into the woman's womb for
implantaton.
 The creation of 'female' sperm and
'male' eggs
• If this technological breakthrough ever comes
about, it is suggested that such artificial gametes
could offer the possibility of genetic reproduction
to people in same sex relationships.
• In fact artificial gametes offer people to have
genetically related offspring regardless of age,
gender, relationship status or sexuality. Women
could use the technique to produce eggs even
after having gone though the menopause.
• Individuals who cannot find reproductive
partners could even use artificial gamete
technology to create complementary gametes
from their own bodies to fertilize their 'natural'
sperm or eggs.
 Applications of ART
• Endanger species
• Assisted reproductive technology, allows in
vitro fertilized (IVFed) embryos for
preimplantation genetic screening (PGS)
evaluation. The is not used to look for a
specific disease but a technique to identify
embryos at risk.
 The End

Thank you!!