1. Dr. ZEINAB ABOTALIB MRCOG, DGO, Consultant Obs/Gyna Infertility & IVF
Assisted Conception
Dr. ZEINAB ABOTALIB MRCOG, DGO,
Associate Professor &
Consultant Obs/Gyna Infertility & IVF

2. Assisted Conception
IUI: intrauterine insemination
IVF: in vitro fertilization
ICSI: intracytoplasmic sperm injection
GIFT: gamete intrafallopian transfer
ZIFT: zygote intrafallopian transfer
PESA: percutaneous epididymal sperm aspiration

3. Assisted Conception
ET: embryo transfer
TESE: testicular sperm extraction
SUZI: subzonal sperm injection
PGD: preimplantation genetic diagnosis

4. Assisted Conception
Objective
To bring sperm and oocyte close to each other to promote chances of fertilization and, ultimately, achieve a pregnancy

5. Assisted Conception
Main types:
IUI: intrauterine insemination
IVF: in vitro fertilization
ICSI: intracytoplasmic sperm injection

6. Assisted Conception
Required procedures
Superovulation
Sperm preparation
Assisted fertilization

7. Superovulation
Hormonal manipulation to enhance ovulation and release multiple oocytes during ovulatory cycle

8. Superovulation Drugs used: Human menopausal gonadotropin
Taken from urine of postmenopausal women
Follicle stimulating hormone (FSH) and luteinizing hormone (LH) activity
Recombinant FSH
Recombinant LH

9. Superovulation- protocol
Gonadotropin for 9-11 days
Monitoring follicular development by transvaginal ultrasound
Follicles mm in diameter
10,000 IU hCG
Oocytes maturation
Ovulation

11. Sperm Preparation Select PMNS Remove seminal plasma, WBC, and bacteria
Sperm capacitation
Coating of sperm with seminal plasma proteins
Allow sperm to become fertile
In vivo or in test tube

13. Intrauterine insemination
Sperm sample deposited in uterus just before release of an oocyte (s) in a natural or stimulated cycle
Soft catheter
Give hCG at injection or up to 24 hrs later
Sperm volume: ml
Pregnancy rates
Around 15% per cycle

14. Gamete intrafallopian transfer
Laparoscopic technique in which oocyte and sperm placed in fallopian tube, allowing in vivo fertilization
Procedure
Superovulation
US guided transvaginal oocyte retrieval
mil sperm with 2-3 oocytes

16. In vitro fertilization - IVF
Taking oocytes from woman
Fertilizing them in lab with her partner's sperm
Transferring resulting embryos back to her uterus 3 or 5 days later

17. IVF Procedure Superovulation Insemination Embryo transfer
Luteal support

18. IVF - Superovulation Gonadotropin stimulation
Monitoring follicular development
US guided transvaginal oocyte retrieval
Oocyte fertilization with sperm

21. IVF - Insemination Containers used
Test tubes, Petri dishes, multi-well dishes
Each oocyte inseminated with mil PMNS
Fertilization detected hrs later by presence of
2 pronuclei in oocyte cytoplasm
2 polar bodies in perivitelline space

22. IVF - Insemination
Syngamy (combination of maternal and paternal pronuclei 24 hrs after insemination
Further cleavages occur at 24 hr intervals

26. IVF - Embryo transfer
Embryos transferred to uterus on 2nd or 3rd day after in vitro insemination
4-8 cells embryos
2-3 embryos transferred in 20 µl of culture fluid
Transabdominal US to see fluid placed in uterus
Cryopreserve excess embryos

28. IVF - Luteal support
Progesterone (P4) necessary for pregnancy maintenance
Premature luteolysis in some superovulatory regimens
P4 supplementation until menses occur or woman has positive pregnancy test

29. Intracytoplasmic sperm injection - ICSI
Injection of single sperm into single oocyte in order to get fertilization
Procedure
Superovulation
US guided transvaginal oocyte retrieval
IVF
Oocytes injected with sperm using special microscopes, needles and micromanipulation equipment

35. ICSI - Indications
Low sperm concentration, motility, abnormal morphology
Antisperm antibodies
Fertilization failure after conventional IVF
Ejaculatory disorders
absence of vas deferens or obstruction of ejaculatory ducts

36. Assisted Hatching Indications Couples having IVF with
Female partner's age over 37
Poor quality embryos
Excessive fragmentation
Slow rates of cell division

37. Assisted Hatching – Procedure
Embryo held with a specialized holding pipette
A needle used to expel an acidic solution against ZP
A small hole made in ZP
Embryo washed and put back in culture in incubator
ET shortly after hatching procedure
Hope for the best

41. Further Advances And Uses Of Assisted Conception Technology
Cryopreservation of
Sperm
Embryo
Oocyte
Ovarian tissue
Growth of human follicles and oocytes in vitro
In vitro maturation and transplantation of human spermatozoa

42. Assisted Reproductive Technology (ART)
Dr. Abdelsalam Talafha
American Board Certified, Comparative Veterinary Obstetrics and Gynecology

43. ART
Infertility
Inability to conceive after 1 year of unprotected and regular sexual intercourse
Primary infertility
Couples have never had children
Secondary infertility
Couples initiated conception in the past and then had difficulty

44. ART Infertility Female partner: 35% Male partner: 35%
Both partners: 20%
Unknown cause: 10%
Infertility more common with increasing age

45. ART USA women infertility rate Ages 20-24: 4.1% Ages 25-29: 5.5%
80% of infertility cases can be diagnosed
85% of cases can be successfully treated

46. ART Female infertility Disorders of ovulation: 27%
Fallopian tube disorders: 22%
Pelvic adhesions: 12%
Endometriosis: 5% to 15%
Hyperprolactinemia: 7%

47. ART Male infertility Abnormal semen parameters
Count, motility, morphology
Infertility treatment
Correcting underlying abnormality
ART

48. ART
Main techniques
IUI
IVF - embryo transfer
ICSI
Assisted hatching

49. Preimplantation Genetic Diagnosis (PGD)
Identify genetic conditions in embryo before ET
Hemophilia
Cystic fibrosis
Aneuploidy

50. PGD Hemophilia Hereditary bleeding disorder
Absence of a blood protein essential for clotting
Types A: lack of factor VIII
Type B: lack of factor IX

51. PGD Cystic fibrosis Genetic disease
Defective gene causes body to produce abnormally thick, sticky mucus that obstruct
Lungs
Pancreas

52. PGD
Aneuploidy
Having less than or more than normal diploid number of chromosomes
Monosomy
Trisomy
Triploidy

53. PGD Performed with IVF 8-cell stage (3 days old) embryo biopsy
Obtain 1-2 blastomeres for genetic

55. Three day old embryos

60. PGD
Genetic analysis
Multicolor fluorescence in situ hybridization (FISH)
Polymerase chain reaction (PCR)

61. FISH Detects Number of chromosomes Sexing embryos
Sex chromosome aneuploidy
Whole-chromosome paints for detection of rearrangements and identification of marker chromosomes
Analyze polar bodies

62. FISH
Fuorescent probes that bind to specific chromosomes are labeled with biotin for detection by specific fluorochrome-conjugated antibodies under a fluorescent microscope

63. Interphase nucleus X Y

64. Triploid
Normal Diploid
Tetraploid
Missing chromosomes

65. PGD
Double trisomy
Monosomy