Assisted reproductive technology (art)

i. Intrauterine insemination (IUI).
ii. In vitro fertilization and embryo transfer (IVF and ET).
iii. Gamete intra-fallopian transfer (GIFT).
iv. Zygote intra-fallopian transfer (ZIPT).
v. Intra-vaginal culture (IVC).
vi. Cytoplasmic transfer (CT).

1. Assisted reproductive technology (ART) Dr. P. Nithiya Assistant professor Department of Botany Seethalakshmi Ramaswami college Tiruchirappalli

2. Assisted reproductive technology (ART) Assisted reproductive technology (ART) is used to treat  infertility . It includes fertility treatments that handle both a woman's egg and a man's sperm. It works by removing eggs from a woman's body. The eggs are then mixed with sperm to make embryos . The embryos are then put back in the woman's body. In vitro fertilization (IVF) is the most common and effective type of ART.

3. Assisted reproductive technology (ART) ART procedures sometimes use donor eggs, donor sperm, or previously frozen embryos. It may also involve a surrogate or gestational carrier. A surrogate is a woman who becomes pregnant with sperm from the male partner of the couple. A gestational carrier becomes pregnant with an egg from the female partner and the sperm from the male partner. The most common complication of ART is a  multiple pregnancy . It can be prevented or minimized by limiting the number of embryos that are put into the woman's body.

4. Techniques involved in ART Assisted Reproductive Technology (ART) includes i . Intrauterine insemination (IUI). ii. In vitro fertilization and embryo transfer (IVF and ET). iii. Gamete intra-fallopian transfer (GIFT). iv. Zygote intra-fallopian transfer (ZIPT). v. Intra-vaginal culture (IVC). vi. Cytoplasmic transfer (CT). vii. Micromanipulation (Intra-cytoplasmic sperm injection (ICSI), sub-zonal insertion (SUZI). viii. Cryopreservation. ix. Assisted hatching (AH).

5. Techniques we are going to study i . Intrauterine insemination (IUI). ii. In vitro fertilization and embryo transfer (IVF and ET). iii. Gamete intra-fallopian transfer (GIFT). iv. Zygote intra-fallopian transfer (ZIPT). v. Intra-vaginal culture (IVC). vi. Cytoplasmic transfer (CT ).

6. I. INTRAUTERINE INSEMINATION (IUI) The infertile women (due to endometriosis, idiopathic infertility) without blockage or damage to fallopian tubes can be effectively treated by intrauterine insemination . The women with adequate ovulation and below the age of 40 years are considered for IUI. The women are usually super-ovulated by administering gonadotrophins . This results in multiple egg development. The IUI is timed to coincide with ovulation . The semen is washed and the highly motile sperms are separated.

7. I. INTRAUTERINE INSEMINATION (IUI) By using a thin and soft catheter, the sperms are placed either in the cervix or in uterine cavity. The women subjects are advised to remain lying down for about 15-30 minutes following IUI.  Insemination should be carefully timed for good success. If it is done, a little before the expected time of ovulation, the chances for fertilization are much higher. IUI is usually successful in the first 3-4 attempts. In any case, this approach is not recommended for more than a maximum of 6 ovulation cycles. The success rates of IUI vary considerably and are in the range of 15-30%.

8. I. INTRAUTERINE INSEMINATION (IUI)

9. In vitro Fertilization In vitro fertilization broadly deals with the removal of eggs from a women, fertilizing them in the laboratory, and then transferring the fertilized eggs (zygotes) into the uterus a few days later.

10. Indications for IVF: Infertility due to the following causes may be considered for IVF. i . Failed ovulation induction ii. Tubal diseases iii. Cervical hostility iv. Endometriosis v. Idiopathic infertility (in men and women).

11. Ideal Subjects for IVF : Although it is not always possible to have a choice in the selection of subjects, the following criteria are preferred. i . Woman below 35 years. ii. Presence of at least one functional ovary. iii. Husband with normal motile sperm count (i.e. normal seminogram ). iv. The couple must be negative for HIV and hepatitis.

12. Methodology of IVF: The in vitro fertilization broadly involves the following steps. 1. Induction of superovulation. 2. Monitoring of ovarian response. 3. Oocyte retrieval. 4. Fertilization in vitro. 5. Embryo transfer.

13. IVF

14. Induction of Superovulation : It is well known that the success rate IVF is much higher when more embryos (3-5) are transferred. This is possible only with controlled ovarian hyper-stimulation (COH). The other advantages of COH include improvement in the quality of oocyte, control of ovulation timing, besides overcoming the ovulatory dysfunction. The following drug regimens are in use to induce superovulation. i . Clomiphene citrate (CC). ii. CC + human menopausal gonadotrophin ( hMG ). iii. CC + follicle stimulating hormone (FSH ).

15. Induction of Superovulation: iv. Human menopausal gonadotrophin . v. Follicle stimulating hormone. vi. Gonadotrophin releasing hormone agonists ( GnRHa ) + hMG (or FSH). It is now common to use GnRH agonists to induce ovulation. These compounds act through a process called down regulation of the physiologic hypothalamic- pituitary-ovarian feedback mechanism to effectively suppress spontaneous ovulation.

16. Monitoring of Ovarian Response: The follicular growth or ovarian response can be monitored by increase in serum estradiol level, increase in follicular diameter and thickening of endometrial bed.

17. Oocyte Retrieval: The most common method for oocyte retrieval is carried out through vaginal route under ultrasound guidance. This method is simple and less invasive, and can be performed with analgesics only . It is easy to recognize the oocyte as a single cell surrounded by a mass of cumulus cells. The recovered oocytes are maintained in vitro culture for 4-6 hours.

18. Fertilization in Vitro : The semen specimens are collected (just prior to oocyte retrival ) via masturbation, processed, and incubated in protein-supplemented media for 3-4 hours prior to fertilization. The incubation results in sperm capacitation.

19. Embryo Transfer: Embryo at a stage between pronuclei and blastocyst stage are transferred. Conventionally , 4- 8 cell stage embryos are transferred between 48-60 hours following insemination. The transfer procedure is carried out by use of a catheter. Not more than three embryos are transferred (per cycle) to minimize multiple pregnancies. However , in the women above the age of 40 years, higher number of embryo may be transferred.

20. Note : Excess oocytes and embryos are cryopreserved for further use. This will reduce the cost, besides the risk of ovarian hyper stimulation

21. Louise Brown was born in UK in 1978

22. Kanupriya alias Durga was born in Kolkata on 3rd October 1978.

23. The success rate of IVF is rather low due to the following reasons: The success rate of IVF is rather low due to the following reasons: i . Increased risk of abortion ii. Multiple pregnancy iii. Ectopic pregnancy iv. Low birth weight baby v. Premature delivery.

24. Thank You

25. Gamete Intra-Fallopian Transfer (GIFT ). Dr. P. Nithiya Assistant professor Department of Botany Seethalakshmi Ramaswami college Tiruchirappalli

26. Gamete intrafallopian transfer (GIFT) Gamete intrafallopian transfer (GIFT) was first developed in 1984 as part of further studies into  assisted reproductive technologies  (ART). 

27. Gamete intra-fallopian transfer Gamete intra-fallopian transfer involves the transfer of both sperm and unfertilized oocyte into the fallopian tube. This allows the fertilization to naturally occur in vivo. The prerequisite for GIFT procedure is that the woman should have at least one normal fallopian tube.

28. Gamete intra-fallopian transfer The induction of ovulation and the monitoring procedures for GIFT are almost the same as described for IVF. A couple of hours prior to oocyte retrieval, semen specimens are collected. Two oocytes along with 2-5 lakhs motile sperms for each fallopian tube are placed in a plastic tube container. It is then inserted (by laparoscopy) 4 cm into the distal end of the fallopian tube, and the oocyte sperm combination is injected.

29. Gamete intra-fallopian transfer The overall pregnancy rate is as high as 30- 40%. The take home baby rate is about 25%. This is much higher when compared to IVF. But the major limitation is the requirement of laparoscopy (a major surgical procedure) to transfer oocytes and sperms into the fallopian tubes.

30. Gamete intra-fallopian transfer The overall pregnancy rate is as high as 30- 40%. The take home baby rate is about 25%. This is much higher when compared to IVF. But the major limitation is the requirement of laparoscopy (a major surgical procedure) to transfer oocytes and sperms into the fallopian tubes.

31. Gamete intra-fallopian transfer Like IVF, GIFT involves removing eggs from your ovaries, mixing them with her partner's sperm and returning them to her body. Unlike IVF, after the eggs and sperm (the gamets ) are mixed, they are immediately transferred to your fallopian tubes rather than to a laboratory dish.

32. Gamete intra-fallopian transfer With GIFT, fertilization happens in your fallopian tubes rather than an incubator . Just as in a natural pregnancy, the fertilized egg(s) moves down your fallopian tubes and develops into an embryo, ultimately implanting into your uterus . Because the eggs are fertilized inside the body, you will not need to discard, donate or  store extra embryos .

33. On the down side, GIFT also means that: While doctor may evaluate and choose the healthiest eggs, Patient has no way of knowing if the eggs will become fertilized . In IVF, your eggs are placed in the uterus  after  fertilization has occurred. You will have two different surgical procedures: egg retrieval (as in all ART procedures), and then laparoscopic surgery to place the eggs and sperm in to the fallopian tubes.

34. Problem associated with GIFT Postoperative bladder infection Skin irritation Hematomas (blood-filled bruising) of the abdominal wall Pelvic or abdominal infections

35. Zygote Intra-Fallopian Transfer (ZIFT ) Dr. P. Nithiya Assistant professor Department of Botany Seethalakshmi Ramaswami college Tiruchirappalli

36. Zygote Intra-Fallopian Transfer (ZIFT) ZIFT is suitable when the infertility lies in men, or in case of failure of GIFT. The wife’s oocytes are exposed to her husband’s sperms in the laboratory. The fertilized eggs (zygotes) within 24 hours are transferred to the fallopian tube by using laparoscopy. ZIPT has an advantage over GIFT with male factor infertility. Further , it can be known whether the wife’s oocytes have been fertilized by her husbands’ sperms.

37. Zygote Intra-Fallopian Transfer (ZIFT) A technique in which a woman's egg is fertilized outside the body, then implanted in one of her fallopian tubes. This technique is one of the methods used to overcome infertility, the inability of couples to produce offspring on their own. First , the egg and the male sperm needed to fertilize it are harvested.

38. Zygote Intra-Fallopian Transfer (ZIFT) Then the egg and the sperm are united in a petri dish, a multi-purpose glass or plastic container with a lid. If all goes well, the sperm fertilizes the egg, and the physicians then implant it in a fallopian tube. From there, nature takes its course, and the egg eventually is deposited by the fallopian tube into the uterus (womb) for development.

39. Zygote Intra-Fallopian Transfer (ZIFT) A zygote is the combined cell resulting from the union of sperm and egg. A zygote develops into an embryo. An embryo, a mass of cells with no recognizable human features, begins formation of a human body. After about seven or eight weeks, the embryo exhibits recognizable features such as a mouth and ears.

40. Zygote Intra-Fallopian Transfer (ZIFT) At this stage, the developing human becomes known as a fetus. The word "zygote" is derived from the Greek word " zygon " (yoke). The term " intrafallopian " means "inside the fallopian tubes." ("Intra," a Latin word, means "within" or "inside.") Thus, the term "zygote intrafallopian transfer" refers to the transfer of a zygote into a fallopian tube.

41. Zygote Intra-Fallopian Transfer (ZIFT)

42. Intra Vaginal Culture (IVC ) Dr. P. Nithiya Assistant professor Department of Botany Seethalakshmi Ramaswami college Tiruchirappalli

43. Intra Vaginal Culture (IVC ): The body’s own environment is appropriately utilized in intra-vaginal culture. The retrieved oocytes and sperms are placed in a culture medium inside a sealed container . This is inserted into the vagina. The container is held by a vaginal diaphragm . Thus, the oocytes and sperms are maintained at the normal body temperature (in contrast to any incubator in the laboratory).

44. Intra Vaginal Culture (IVC ) Two to 3 three days later, the container is opened, and the fertilized and dividing zygotes are transferred into the uterus. This procedure appears simple, but the success rate is very low. Only a few centers practice this.

45. Cytoplasmic Transfer (CT ) Dr. P. Nithiya Assistant professor Department of Botany Seethalakshmi Ramaswami college Tiruchirappalli

46. Cytoplasmic Transfer (CT): Cytoplasm includes many things, the most important being mitochondria which provide energy to the cell. It is possible that deficiency in the mitochondria may leave the oocyte without the necessary power for cell division, after fertilization . This may result in abnormal cell division and poor development of embryo.   It is therefore logical to think of the transfer of cytoplasm from a donor (with active mitochondria) into the oocyte of a woman. The advantage with cytoplasmic transfer is that the mother’s own genetic material is passed on to the offspring.

47. Two methods of cytoplasmic transfer have been developed:   1 . Transfer of a small amount of cytoplasm by a tiny needle from a donor to a recipient oocyte.   2 . Transfer of a large amount of cytoplasm which is fused with the recipient’s cytoplasm by applying electricity. The procedure of cytoplasmic transfer is tedious and technically difficult, besides the cost factor. At least two viable pregnancies have been so far reported in literature by this approach.

48. Thank you