in vitro fertilization
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  • 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.
  • 5.  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
  • 6. 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
  • 7. GENERAL PROTOCOL  Patient selection  Manipulation of mensural cycle; superovulation  Oocyte retrieval  Preparation of semen sample  IVF treatment  Embryo transfer  Cryopresevation
  • 8. 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.
  • 9. 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.
  • 10. 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.
  • 11. 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.
  • 12.  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
  • 13.  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)
  • 14.  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.
  • 15.  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)
  • 16.  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.
  • 17. 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.
  • 18.  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)
  • 19.  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.
  • 20. 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.
  • 21. 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.
  • 22. 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.
  • 23. 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