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Andrology and IVF Laboratory Services. Charles L. Bormann, PhD IVF/Andrology Laboratory Supervisor. Clinical Laboratory Improvement Amendments (CLIA) Food and Drug Administration (FDA) College of American Pathologists (CAP) Society for Assisted Reproductive Technology (SART).

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Andrology and IVF Laboratory Services


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    1. Andrology and IVF Laboratory Services Charles L. Bormann, PhD IVF/Andrology Laboratory Supervisor

    2. Clinical Laboratory Improvement Amendments (CLIA) Food and Drug Administration (FDA) College of American Pathologists (CAP) Society for Assisted Reproductive Technology (SART) Mark P.D. Dow PhD, HCLD (Laboratory Director) Charles L. Bormann MS, PhD (Laboratory Supervisor) Lynn Boehnlein BS, MT,TS (Senior Embryologist) Elizabeth Schleinz BS, MT,TS (Senior Embryologist) Laboratory Regulations

    3. Semen Analysis • Abstain 2-3 days prior to analysis • Collect in an approved sterile cup or condom • Do not use lubricants for collection • Sample processed within 1 hour of collection

    4. World Health Organization Parameters Volume >2ml pH 7.2-8.0 Sperm Concentration >20 million/ml Sperm Motility >50% Progression/Speed ≤3 Sperm Viability ≥50% Morphology (Krueger) ≥14% Round Cells ≤5/hpf DNA Fragmentation ≤30% Viscosity

    5. Sperm Motility *Motility *Forward Progression *Velocity

    6. Sperm Morphology

    7. Sperm DNA Fragmentation <30% DNA Fragmentation

    8. Sperm DNA Fragmentation Density Gradient Swim-Up Density Gradient Spin Wash

    9. Sperm Viability Tests • Viability Stain • Pentoxifylline • Hypo-Osmotic Swelling (HOS)

    10. Donor Sperm • Purpose: Offer a chance at pregnancy to those without a male partner, or if male partner’s sperm are absent, unable to fertilize, carry a genetic disease, etc

    11. Donor Sperm Photos and personal information also available

    12. Inter-Uterine Insemination (IUI) Semen • Remove seminal plasma • Remove debris (WBC, dead sperm) • Enrich highly motile sperm population Wash DGS Centrifuge, remove supernatant Centrifuge, remove top layers Wash Wash Count/Assess Reconstitute Load catheter

    13. Intracytoplasmic Sperm Injection (ICSI)

    14. Fertilization ~ 18h post insemination Female Pronucleus Male Pronucleus 2 1 3 1 1 4 2 6 5 3

    15. Embryo Development

    16. In Vitro Culture The in vitro environment was designed to mimic mother nature

    17. Embryo Development 4-Cell 8-Cell 2-Cell Days 1-3 Hatching Blastocyst Blastocyst Morula Days 4-6

    18. Day 3 vs. Day 5 Embryo Transfer 8AA 8AA 8AA 8AA 4AA 4AB 2CB 3BB

    19. Preimplantation Genetic Diagnosis • Recurrent Miscarriage • Unsuccessful IVF Cycles • Unexplained Infertility • Advanced Maternal Age • Male Factor Infertility • Inherited Genetic Disorders

    20. Preimplantation Genetic Diagnosis (PGD) Day 3 Embryos (8-Cell) Embryo Culture Days 1-3 Blastomere Biopsy Day 3 Day 5-6 Blastocyst Culture, Embryo Transfer and Possible Freeze

    21. Assisted Hatching Application • Abnormally thick zona pellucida • Frozen/thawed embryos/oocytes • Prior failed IVF • >37 yrs of age • Elevated FSH

    22. Fertility Preservation

    23. Sperm Freezing • Frozen Indefinitely • Simple to Freeze • No documented risks from using frozen sperm

    24. Egg Freezing -Considered Experimental -Option for the preservation of fertility in women with malignant diseases -Not ideal for women wishing to delay childbirth

    25. Embryo Freezing -Preserve additional embryos for future cycles -Embryos frozen indefinitely

    26. 2007 SART National Summary: Age ~5-10% Lower with Frozen Embryos

    27. Semen Analysis Identification • Identification • Labeling • Chain of Custody • Isolation • Verification Name: John Doe DOB: March, 23, 2010 MR#: 87654321

    28. IVF/ICSI Identification • Identification • Labeling • Chain of Custody • Isolation • Verification

    29. Doctors That Are Happy To See You