1. By,
DAMARIS BENNY DANIEL
II Msc. ZOOLOGY

2. INTRODUCTION
 Medically assisted human reproductive technologies are a
group of high tech treatment methods used to combat infertility.
 One of the greatest advances in reproductive medicine
 Techniques include
 Intrauterine Insemination (IUI)
 In Vitro Fertilization and
Embryo transfer (IVF – ET)
 Gametic Intra-Fallopian Transfer (GIFT)
 Zygotic Intra-Fallopian Transfer (ZIFT)
 Tubal embryo stage transfer (TET)
 Intracytoplasmic sperm injection (ICSI)

3. CAUSES OF INFERTILITY
 In males
 Oligospermia – reduced conc. of sperm in semen.
 Azoospermia – Total lack or very low conc. of motile sperm
 In females
 Tubal infertility – non functional fallopian tube
 Non functional ovaries
 Non functional uterus
 Endometriosis - Endometrial-like cells in areas outside the
uterus
 Idiopathic infertility – reason unknown.

4. INTRA UTERINE INSEMINATION (IUI)
 Women (with adequate
ovulation & below 40yrs)
without damage to fallopian
tube can be treated with IUI.
 Women superovulated by
gonadotrophins – multiple
egg dev.
 IUI is timed to coincide with
ovulation
 Using a thin soft catheter,
sperms are placed either in
the cervix or in the utrine
cavity.

6.  ADVANTAGES
 This procedure can be an effective solution for men with a low
sperm count or poor sperm motility, where the sperm can't
make the long journey to the egg.
 Low cost compared to other ART .
 DISADVANTAGES
 Timing in the process of insemination is important.
 Low success rate, results are only 10-20 %
 Fallopian tubes must be unobstructed

7. IN VITRO FERTILIZATION
 IVF broadly deals with the removal of eggs from a women,
fertilizing them in the laboratory and then transferring the
fertilized eggs into uterus a few days later.
 Infertility due to the following causes may be considered for IVF
 Failed ovulation induction
 Tubal diseases
 Endometriosis
 Idiopathic infertility

8. GENERAL PROTOCOL
 Patient selection
 Manipulation of mensural
cycle; superovulation
 Oocyte retrieval
 Preparation of semen
sample
 IVF treatment
 Embryo transfer
 Cryopresevation

9. 1) PATIENT SELECTION
Criteria
 Woman below 35yrs
 Presence of atleast one functional ovary
 Husband with normal motile sperm count
 The couple must be negative for HIV and hepatitis.
2) INDUCTION OF SUPEROVULATION
 1 week before next menstural period – leuprolide injection –
prevent premature release of egg.
 Within 2 week of onset of menstural period – FSH injection –
stimulates maturation of multiple eggs.
 Then a single injection of HCG (Human Chorionic Gonadotropin)
is given – triggers final stage of egg maturation. Ovulation will
occur between 24–36 hours after the HCG injection.

10. 3) OOCYTE RETRIEVAL
 Eggs are retreived after 36 hrs of HCG injection.
 Most common - through vaginal route under ultrasound
guidance.
 Needle enters the ovarian follicle and aspirate the fluid from
them
 It is examined under microscope to identify eggs.
4) PREPARATION OF SEMEN SAMPLE
 Semen collected from the partner.
 Processed and incubated in protein supplemented media for
3-4 hrs – rslts in sperm capacitation.

11. 5) IVF TREATMENT
 Capacited sperms placed in culture with a single oocyte in a
petridish.
 Signs of fertilization – presence of 2 pronuclei
6) EMBRYO TRNSFER
 4 – 8 cell stage embryos are transferred
 Transferred with the help of a catheter.
 Catheter is passed through the cervical canal and embryos are
released to the top part of the uterus.
 Not more than 3 embryos are transferred.

12. 7) CRYOPRESERVATION
 Preservation in frozen state is regarded
as cryopreservation.
 Semen, fertilized eggs and embryos can be cryopreserved.
 Human embryos have been successfully preserved in the
presence of cryoprotectant like 1, 2 propanediol or dimethyl
sulphoxide or glycerol.
 It was stored at -196OC under liquid nitrogen.
 At appropriate time, the embryos are thawed and is transferred
to uterus.

14.  ADVANTAGES
 Fertilization is confirmed before implantation can occur.
 Gives women with damaged oviducts, the opportunity to
carry their own fetus.
 DISADVANTAGES
 Implantation in the uterus does not always occur.
 Higher risk of twins or triplets, which also increases the risk
of complications and miscarriages.
 Side effects associated with the fertility medication
 Higher risk of ectopic pregnancy, especially in women that
have had previous problems with their oviducts

15.  It involves the transfer of both sperm and unfertilized oocyte into
the fallopian tube.
 This allows the fertilization to naturally occur in vivo.
 Two oocyte along with 2 – 5 lakhs motile sperms are placed in a
plastic tube container.
 Then oocyte sperm combination is injected 4cm into the distal
end of fallopian tube.
GAMETE INTRA FALLOPIAN TRANSFER (GIFT)

16.  ADVANTAGES
 There is no much human intervention in the actual
fertilization of the eggs.
 Because fertilisation takes place within the fallopian tube,
GIFT offers an option for people whose religious beliefs
prohibit conception outside the body.
 DISADVANTAGES
 Can be performed only if woman have atleast one normal
fallopian tube.
 GIFT does not allow for visual confirmation of fertilisation.
 GIFT involves a laproscopic surgery.

17.  ZIFT combines aspects of both IVF
and GIFT.
 Fertilization takes place outside
the uterus and placed into the
fallopian tubes
 Protocols for ovarian stimulation
are similar to those used for IVF
and GIFT.
 Eggs are collected and fertilized
by the partner’s sperm in the
laboratory.
 The zygote is transferred to the
fallopian tube within 24hrs, when
it is at 1 cell stage.
ZYGOTE INTRA FALLOPIAN TRANSFER (ZIFT)

18.  ADVANTAGES
 Fertilization can be confirmed before they are implanted into
the fallopian tube.
 Allows a developing embryo to travel into the uterus on its
own, which may be important to those who wish their baby to
develop as naturally as possible
 DISADVANTAGES
 Can be performed only if woman have atleast one normal
fallopian tube.
 It is more expensive than GIFT.
 ZIFT involves a laproscopic surgery.

19. TUBAL EMBRYO STAGE TRANSFER (TET)
 It combines IVF with tubal transfer
 Embryos are placed into the women’s fallopian tube.
 The embryos are transferred back into the woman 2 days after
fertilisation. This is at the ‘2 cell or 4 cell’ stage.
 TET allows embryos to make their way to the uterus for
implantation.
 Its advantage over ZIFT is that it allows for the assessment of
fertilization and embryo quality.
 Success rate higher than ZIFT.

21.  Sperm is injected directly into the eggs in a laboratory.
 Used if infertility originates from the male such as:
 Low numbers of sperm
 Low sperm motility
 Single spermatozoan is directly injected into the cytoplasm of
the oocyte through the micropuncture of zona pellucida.
INTRA CYTOPLASMIC SPERM INJECTION (ICSI)

22.  ADVANTAGES
 Can be useful when very low numbers of motile sperm are
present and when there are problems with sperm binding and
penetration.
 DISADVANTAGES
 Altering the nature’s selection process for sperm can lead to
an increase risk of developmental and health issues for ICSI
children, as well as a higher risk of miscarriage because of the
poorer genetic material involved.

24. NEGETIVE ASPECTS OF ART
 Due to administration of hormones and drugs, ovarian
hyperstimulation syndrome (OHSS) can occur.
 Risks associated with pregnancy
 Multiple pregnancy
 Increased risk of premature labor etc..
 Can cause premature menopause.
 Increased risk of ovarian cancer, atleast by 3 times when
compared to normal women.

25. CONCLUSION
 ART has been the answer for many childless couples, resulting in
successful pregnancies and childbirth.
 Today there is a range of infertile treatment that aims to ensure a
healthy sustainable pregnancy.
 But there are still risks, stress and high cost associated with ART.
 With the advent of new technologies, it is hoped that these
shortcomings would be overcome in the near future and every
infertile couple would have the previlage of parethood.

26. RECENT TRENDS
 Innovative design of IVF equipment (PLoS ONE, june 2012)
 A novel system for processing embryos during IVF treatment has
been shown to significantly improve the chances of pregnancy by
27%.
 Pioneered by a Newcastle team of fertility experts within the
University and NHS, the innovative design of interlinked
incubators provides a totally enclosed and controlled
environment within which every step of the IVF process can be
performed.

27. BIBLIOGRAPHY
 Satyanarayana U, Biotechnology (2010), 1st edition, Books and
allied (P) Ltd, Kolkata.
 Joseph. C. Daniel, Methods in mammalian embryology, Ist edition,
W.H Freeman and company, San Fransisco
 www. sart. org/publications/details.aspx? Id = 1522
 www.youtube.com/
 http://www.ncl.ac.uk/press.office/press.release/item/increased-
fertility-rate-for-ivf-patients-achieved-by-new-equipment-design