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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 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) 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
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
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
Sperm Motility *Motility *Forward Progression *Velocity
Sperm DNA Fragmentation <30% DNA Fragmentation
Sperm DNA Fragmentation Density Gradient Swim-Up Density Gradient Spin Wash
Sperm Viability Tests • Viability Stain • Pentoxifylline • Hypo-Osmotic Swelling (HOS)
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
Donor Sperm Photos and personal information also available
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
Fertilization ~ 18h post insemination Female Pronucleus Male Pronucleus 2 1 3 1 1 4 2 6 5 3
In Vitro Culture The in vitro environment was designed to mimic mother nature
Embryo Development 4-Cell 8-Cell 2-Cell Days 1-3 Hatching Blastocyst Blastocyst Morula Days 4-6
Day 3 vs. Day 5 Embryo Transfer 8AA 8AA 8AA 8AA 4AA 4AB 2CB 3BB
Preimplantation Genetic Diagnosis • Recurrent Miscarriage • Unsuccessful IVF Cycles • Unexplained Infertility • Advanced Maternal Age • Male Factor Infertility • Inherited Genetic Disorders
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
Assisted Hatching Application • Abnormally thick zona pellucida • Frozen/thawed embryos/oocytes • Prior failed IVF • >37 yrs of age • Elevated FSH
Sperm Freezing • Frozen Indefinitely • Simple to Freeze • No documented risks from using frozen sperm
Egg Freezing -Considered Experimental -Option for the preservation of fertility in women with malignant diseases -Not ideal for women wishing to delay childbirth
Embryo Freezing -Preserve additional embryos for future cycles -Embryos frozen indefinitely
2007 SART National Summary: Age ~5-10% Lower with Frozen Embryos
Semen Analysis Identification • Identification • Labeling • Chain of Custody • Isolation • Verification Name: John Doe DOB: March, 23, 2010 MR#: 87654321
IVF/ICSI Identification • Identification • Labeling • Chain of Custody • Isolation • Verification