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Explore the history, techniques, and procedures of Assisted Reproductive Technology (ART) used to address infertility issues in couples. Learn about the significant figures and milestones in reproductive chronology and the causes of infertility. Discover the various methods, from superovulation to in vitro fertilization (IVF), aimed at assisting couples in conceiving.
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ART • Infertility • Female partner: 35% • Male partner: 35% • Both partners: 20% • Unknown cause: 10% • Infertility more common with increasing age
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
GREGORY PINCUS • 1934 First in vitro fertilization of rabbit eggs • Criticized as “mad scientist” • Developed oral contraceptives in 1950’s
Dr. John Rock1890-1984 • 1936 First doctor to open a Rhythm clinic in Boston • 1944 First IVF; created four embryos in secret • 1954 Collaborated with Pincus on U.S. clinical trials of the pill
STEPTOE AND EDWARDSBirth of Louise BrownFirst IVF Baby, 1978
REPRODUCTIVE CHRONOLOGY • 1934 Gregory Pincus: First animal IVF • 1944 John Rock: First human IVF • 1960 FDA approves “the pill” • 1978 First IVF baby • 2001 421 U.S. ART clinics with 41,000 live births annually
ART • USA women infertility rate • Ages 20-24: 4.1% • Ages 25-29: 5.5% • Ages 30-34: 9.4% • Ages 35-39: 19.7% • 80% of infertility cases can be diagnosed • 85% of cases can be successfully treated
le tecniche • Inseminazione intrauterina • FIVET • ICSI • Crioconservazione ovocitaria e/o embrionaria • IVM
Title • In realtà ognuna di queste “tecniche” prevede l’intervento di più esperti delle diverse componenti che contribuiscono alla completezza del processo. • Le “tecniche” dovrebbero essere applicate conoscendo il più possibile il quadro patologico a cui si stanno applicando. • Lorem ipsum dolor sit amet, consectetuer adipiscing elit. Vivamus et magna. Fusce sed sem sed magna suscipit egestas. • Lorem ipsum dolor sit amet, consectetuer adipiscing elit. Vivamus et magna. Fusce sed sem sed magna suscipit egestas.
Assisted Conception • Techniques designed to assist couples in their effort to procreate • Reason • Identification and correction of female, male or a combination of both infertility causes
Assisted Conception Objective • To bring sperm and oocyte close to each other to promote chances of fertilization and, ultimately, achieve a pregnancy
Assisted Conception Required procedures • Superovulation • Sperm preparation • Assisted fertilization
Superovulation • Hormonal manipulation to enhance ovulation and release multiple oocytes during ovulatory cycle
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
Superovulation- protocol • Gonadotropin for 9-11 days • Monitoring follicular development by transvaginal ultrasound • Follicles 16 -18 mm in diameter • 10,000 IU hCG • Oocytes maturation • Ovulation
1.Artificial Insemination • Intravaginal insemination. • Intracervical insemination. • Intrauterine insemination. • Intratubal insemination.
Indagini diagnostiche di base • Esame seminale con test di capacitazione • (2 esami a distanza di due mesi e in laboratorio adeguato!!!) Recupero di almeno 5 milioni di spermatozoi mobili prima del trattamento e 1.500.000 dopo trattamento • Esami ormonali, ecografia, tampone vaginale • Isterosalpingografia: molto sensibile in popolazioni a basso rischio, meno sensibile in popolazioni a rischio • Se ISG dubbia o patologica, laparoscopia e cromosalpigoscopia
Sperm Preparation • Select PMNS (Progressively motile normal sperm) • 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
Intrauterine Insemination • Sperm sample deposited in uterus just before release of an oocyte (s) in a natural or stimulated cycle using a Soft catheter • Give hCG at injection or up to 24 hrs later • Sperm volume: 0.2-0.3 ml • Pregnancy rates • Around 15% per cycle
In Vitro Fertilization (IVF) • The first successful IVF case was in 1978 when a British woman delivered a baby called Louise Brown in 28 of July 1978 in Oldam in England by cesarean section. • It had taken nearly a century of research to achieve that.
IVF Indications: • -Women with blocked fallopian tubes • -Ovulation problems • -Seminal problems for the man: sperm transport, motility…… • -Unexplained infertility cases. • -Presence of seminal antibodies in the woman’s body
FIV-ET “classica” - indicazioni • Fattore tubarico – pelvico • Endometriosi moderata – severa • Fallimento iter terapeutico (maschile lieve, PCO, inspiegata)
IVF • Procedure • Superovulation • Insemination • Embryo transfer • Luteal support
IVF What are the steps done in IVF? • 1- Ovarian stimulation • 2- Vaginal ultrasound: • - monitor the number and size of the follicles . • - adjust the dose of the injections accordingly. • 3- HCG Hormone injection is given for final maturation of the eggs. • 4- Oocytes (eggs) retrieval is done 34-37 hours after taking the HCG injection. • 5-The fluid collected from the follicles in special tubes is examined by the embryologist and the eggs are isolated. • 6-The semen sample is taken in the same day and the eggs fertilized either using the conventional IVF or ICSI
FIV-ET “classica” • Pick up: 34 – 36 ore dopo la soministrazione di hcg; preferibile la sedazione profonda • Sala operatoria idonea • Aghi mono o bi-via • Lavaggio del follicolo: ininfluente • Profilassi antibiotica: non utilizzata da molti. Preferibile in casi selezionati: endometriosi severa, pregresse PID • età media 36 aa PR 34%; IR 14%
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FIV-ET “classica” • C’è un trend generalizzato (soprattutto in Italia) ad eseguire meno FIVET e più ICSI • Per nessuna indicazione, tranne i fattori maschili severi, la ICSI ha risultati migliori • Donne over 40? C’è la tendenza a fare una ICSI
FIV-ET “classica” • Idoneità strutture • Contiguità sala operatoria – laboratorio • Qualità degli ambienti • Personale previsto dalle Linee Guida • Regolamenti europei, nazionali, regionali • Programma di crioconservazione: reso obbligatorio dalla legge 40/2004.
IVF - Insemination • Containers used • Test tubes, Petri dishes, multi-well dishes • Each oocyte inseminated with 0.5-1.0 mil PMNS • Fertilization detected 12-20 hrs later by presence of • 2 pronuclei in oocyte cytoplasm • 2 polar bodies in perivitelline space
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
ICSI:Intracytoplasmic Sperm Injection • 1988: prima icsi; 1992 Palermo G. prima gravidanza da icsi • Fattore maschile severo • Anticorpi antispermatozoi (Mar, Immunobead, ecc...) • Azoospermia ostruttiva e secretoria • Pazienti oncologici • Lesioni midollari • Precedenti non fertilizzazioni in FIVET • Precedenti fallimenti FIVET
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
ICSI strumentazione aggiuntiva • Micromanipolatore • Holding pipette e injection pipette • Microscopio invertito a 200 o 400 x • Piano antivibrazioni
Methods For Improving IVF • -Transfer embryos at blastocyst stage • -Culture fertilized ova and early embryos with cells that normally surround the oocyte, so they can provide growth factors. • -Screen early embryos for chromosome abnormalities and implant only those with normal karyotypes.
Assisted Hatching Indications • Couples having IVF with • Female partner's age over 37 • Poor quality embryos • Excessive fragmentation • Slow rates of cell division
4.Assisted Hatching : • Indications: • -Repeated implantation failure in special cases • when the wall of the Embryo is thick