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ASSISTED REPRODUCTION TECHNICS

Learn about various insemination and IVF procedures like AIH and ICSI, including indications, preparation, efficacy, risks, and advantages of these techniques. Discover how IVF works, hormonal stimulation, embryo transfer, and the course of IVF treatment.

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ASSISTED REPRODUCTION TECHNICS

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  1. ASSISTED REPRODUCTION TECHNICS

  2. Inseminations : by husband-AIH, by donor-AID intravaginal-impotention, hypospadiasis, retrograde ejaculation, vaginismus cervical - OAT, cervical defects intrauterine - OAT, negative penetration test, idiopathic sterility

  3. Indications to insemination • idiopathic sterility • congenital defects • retrograde ejaculation • sperm’s hypowolemy • OAT • azoospermia • sexual disorder

  4. Insemination- conditions • non- obstructed Fallopian tubes • monitoring of ovulation • induction of ovulation • bacteriological state of vagine, cervix, sperm • min. 1-5 mln sperm cells with progressive motility in 1 ml of sperm

  5. Preparation of sperm • swim up method • filtration in Percoll gradient TARGET: • separation of sperm cells from sperm plasma • selection and increase number of sperm cells with good morphology and motility • contaminations removal /dead sperm cells, bacterium/ • stimulation of capacitation

  6. Insemination performing • 1-3 times in the cycle /optimum before and after ovulation / • USG monitoring • ovulation induction - Clostilbegyt , HMG • verification /HSG , Echovist-test / • resignation after 6-10 unsuccessful inseminations /classification to laparoscopy or IVF/

  7. Efficacy • the highest : AID retrograde ejaculation cervix defects • the lowest : OAT endometriosis male infertility treated with AIH- 7% of pregnancies pro patient / 2,1% pro cycle / disturbation of ovulation-AIH-29% pregnancies pro patient /11% pro cycle/

  8. AID - conditions • male interfility - azoospermia, examination of urinary sediment after ejaculation, biopsy of testes • OAT after unsuccessful AIH and resignation ICSI • transsexualismus • risk of infections and genetic disorders transmission • multiple, unsuccessful IVF or ICSI

  9. AID technic • frozen sperm • sperm from sperm bank • collection during maximally 6 month • 1-3 times in the cycle

  10. Advantages of AID • patient’s safety • anonymous of donor • accessibility of sperm

  11. IVF - indications • absolute :- absent or inoperable tubal obstruction • relative : - tubal obstruction - periadnexal adhesions - idiopathic infertility - multiple, unsuccessful inseminations - endometriosis - male factor - PCO - immunologic infertility - genetic defects - early menopause - oocyte’s donation

  12. Course of IVF • hormonal stimulation - CC, CC + HMG, GnRHa + HMG (SP, LP, Ultra SP, Ultra LP) • monitoring stimulation - USG, E2 • ovulation indication - HCG (Biogonadyl, Pregnyl, Profasi) • punction • preparation of oocytes, sperm cells • insemination and incubation of oocytes in 5% CO2 amd temp. 370 C • evaluation fertilization after 18 hours (2PN) • embryo transfer after 48 hours in st. 4-8 blastomers • freezing supernumerary embryons • suplementation of luteal phase

  13. Assisted Reproduction Technics • male’s factor conditioned - failures - 60-80% • fertilizations - 20-30% inseminated oocytes • lack of fertilizations - 30% /a group with good reproduction’s potential/

  14. Microassisted Fertilization - MAF • facilitation of syngamy by mechanical or chemical dissection of zona pellucida • injection sperms into perivitelline space • injection single sperm cell into oocyte’s cytoplasm

  15. In Vitro Fertilization - IVF • classic • micromanipulations : ICSI SUZI PZD AZH • ZIFT /PROST/ • TET

  16. Partial Zona Dissection - PZD • make possible fusion of sperm cells with olemma and fertilization • mankaments of method: - high percent of oocytes with polispermic fertilization - high percent of non-fertilized oocytes

  17. Subzonal sperm insertion - SUZI • injection of sperm cells /5-15/ under oocyte’s zona pellucida • sperm cells - after capacitation - in the beginning acrosomal reaction • application: - severe oligoastenozoospermia -preceding IVF procedures - without fertilization pregnancy/cycle - 19%, pregnancy/transfer - 27% Polispermic fertilizations - 50%

  18. Intracytoplasmatic Sperm Injection - ICSI • Preparation of sperm: -separation of sperm cells by centrifugation in Percoll gradient -ejaculate with single sperm cells - washing and centrifugation + multiple swim-up method • Preparation of oocytes: - oocyte’s denudation from corona radiata cells / enzymatic and mechanic method/ - oocyte’ s incubation in the 60 IU/ml hialuronidaze’s solution - aspiration into the pipete (diameter of oocyte) - washing in Earle, BM1 HEPES medium

  19. Intracytoplasmatic Sperm Injection - ICSI • Microscopic assessment of oocyte - untouched structure - first polar body - maturityof oocyte: 80% oocytes - MII 20% oocytes -GV/Germinal Vesicle/ GVBD/Germinal Vesicle Braekdown/ MI /Metafase I/ MI+co- culture with Vero line cells - maturity

  20. Intracytoplasmatic Sperm Injection - ICSI • Microinstruments: - injection pipet - external diameter = 7 um - internal diameter = 5 um - holding pipet - external diameter = 60 um - internal diameter = 20 um

  21. Intracytoplasmatic Sperm Injection - ICSI • Methods: - microscope picture with Hoffman’s contrast -micromanipulators -microdrops: with oocytes, with sperm cells -PVP/poliwinylopirolidon/- slowness of sperm cells’ motility -environmental conditions: temp., pH, mineral oil SPERM CELLS: the best kinetic and morphologic parameters OOCYTES: immobilization, positioning

  22. Intracytoplasmatic Sperm Injection - ICSI • Efficiency: 60-70% fertilizations • Failure: - lack of motility sperm cells - injection sperm cells with round heads - oocytes with cytoplasm degeneration - oocyte lesion during procedure - complete lack of fertilization after ICSI - 3% • Risk: congenital defects - 2,7% , chromosomal anomalies - 0,5%

  23. Micromanipulation • ICSI - the most often • PZD - partial zona dissection • SUZI - subzonal sperm insertion • AZH - assisted zona hatching

  24. Micromanipulation -indications • lowered sperm parameters • < 500 000 motility serm cells in the ejaculate • lack of fertilization in preceding IVF procedures or fertilization lower than 5% cells /right sperm parameters/ • obstruction azoospermia

  25. ICSI - course • identical introduction like in IVF procedure • different preparation of oocytes (cleaning from granulosa cells) • micromanipulator’s introduction 1 sperm cell into cytoplasm of mature oocyte

  26. GIFT - conditions • minimally 1 non-obstruction Fallopian tube and ovary • regular uterine • correct sperm

  27. Others • ZIFT /PROST/ - laparoscopic zygote transfer into ampulla of the uterine tube in 2PN stage • TET - laparoscopic embryo transfer into ampulla of the uterine tube

  28. /Testicular Sperm Extraction - TESETesticular Sperm Aspiration - TESA/ • Conditions azoospermia: -dysfunction of testicular tubules fertilization - 60% pregnancies - 30%

  29. Micro-Epidydymal Sperm Aspiration - MESA • application : azoospermia: - lack of deferent duct - obstruction of deferent ducts

  30. FERTILIZATION OF PRECURSOR CELLS OR IMMATURE SPERM CELLS • spermatide injection • spermatide nucleous injection RISK OF DEVELOPMENTAL ABNORMALITIES

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