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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