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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). - PowerPoint PPT Presentation
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Andrology and IVF Laboratory Services
Charles L. Bormann, PhD
IVF/Andrology Laboratory Supervisor
Laboratory Regulations
• 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)
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
Volume
Viscosity
pH
Sperm Concentration
Sperm Motility
Progression/Speed
Round Cells
World Health Organization
>2ml
7.2-8.0
>20 million/ml
>50%
Morphology (Krueger) ≥14%
Parameters
DNA Fragmentation
Sperm Viability
≤3
≤30%
≤5/hpf
≥50%
Sperm Motility
*Motility
*Forward Progression
*Velocity
Sperm Morphology
Sperm DNA Fragmentation
<30% DNA Fragmentation
Sperm DNA Fragmentation
Density Gradient
Density Gradient
Spin Wash Swim-Up
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)
• Remove seminal plasma
• Remove debris (WBC, dead sperm)
• Enrich highly motile sperm population
Wash DGS
Centrifuge,remove supernatant
Wash Wash
Count/Assess
Reconstitute
Load catheter
Centrifuge,remove top layers
Semen
Intracytoplasmic Sperm Injection (ICSI)
11
23
12
34
56
MalePronucleus
FemalePronucleus
~ 18h post insemination
Fertilization
Embryo Development
In Vitro Culture
The in vitro environment was designed to mimic mother nature
Embryo Development2-Cell 4-Cell 8-Cell
Morula Blastocyst Hatching Blastocyst
Days 1-3
Days 4-6
Day 3 vs. Day 5 Embryo Transfer
2CB 3BB 4AB 4AA
8AA 8AA 8AA 8AA
Preimplantation Genetic Diagnosis
• Recurrent Miscarriage• Unsuccessful IVF Cycles• Unexplained Infertility• Advanced Maternal Age• Male Factor Infertility• Inherited Genetic
Disorders
Preimplantation Genetic Diagnosis (PGD)
Embryo Culture Days 1-3
Blastomere Biopsy Day 3 Day 5-6 Blastocyst Culture, Embryo Transfer and Possible Freeze
Day 3 Embryos (8-Cell)
Assisted Hatching
Application
>37 yrs of age Elevated FSH
Abnormally thick zona pellucida Frozen/thawed embryos/oocytes Prior failed IVF
Fertility Preservation
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 Option for the preservation of fertility in women with malignant diseaseswith malignant diseases-Not ideal for women wishing to delay childbirth-Not ideal for women wishing to delay childbirth
Embryo Freezing-Preserve additional embryos for future cycles
-Embryos frozen indefinitely
Fresh Embryos From Non-Donor Oocytes <35 35-37 38-40 41-42
Number of cycles 38,372 21,707 19,099 8,865
Percentage of cycles resulting in pregnancies
45.8 37.2 28.2 18.5
Percentage of cycles resulting in live births 39.9 30.5 21.0 11.7
Age
2007 SART National Summary:2007 SART National Summary:
~5-10% Lower with Frozen Embryos
Semen Analysis Identification
Name: John Doe
DOB: March, 23, 2010MR#: 87654321
• Identification• Labeling• Chain of Custody• Isolation• Verification
IVF/ICSI Identification• Identification• Labeling• Chain of Custody• Isolation• Verification
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