IVF Lab Procedures

»Egg Retrieval

First steps

• Patients are scheduled for surgery
• Physicians use a long needle to puncture ovarian follicles and collect fluid
• Embryologist looks at the fluid under the microscope and finds the egg from each follicle
• Eggs are washed and placed in a dish

»Sperm / Insemination

Finding the best lookers and swimmers

Husbands are asked to give a sperm sample. To get good fertilization the lab tries to isolate fast swimming sperm with good morphology

The sperm sample is placed on a column of silica beads and centrifuged

The “best” sperm reach the bottom fastest and are collected

Motile sperm and eggs are combined to create an embryo. This step is called “Insemination”

»Intracytoplasmic Sperm Injection (ICSI)

Sometimes nature needs a hand

• When sperm counts or motility are too low
  ICSI is used for the insemination step
• A single motile sperm is visualized under the microscope
• The embryologist picks it up with a glass
  needle
• The sperm is then injected into the egg

»Testicular / Epididymal for ICSI

All you need is one sperm per egg

• Sometimes with severe male factor infertility, sperm needs to be surgically isolated from the testis or epididymis by a urologist
• Sperm isolated from the testis or epididymis are injected into oocytes (ICSI)
• IVF lab personnel often search for hours just to find a dozen sperm for injection
• Pregnancy rates of 35% to 60% have been achieved depending on patient age and sperm quality from the testis or epididymis

»Fertilization

Male and female genetic material combine

• Eggs are examined the day after retrieval for fertilization
• The presence of two nuclei in the center of the egg is a sign that
  normal fertilization occurred
• The fertilized eggs are moved to a new dish
  and watched every day for cell division

»Embryo Cleavage

Embryo divides each day

The embryo should divide each day going from a single cell to over 100 cells by the time it attaches to the uterine wall at the blastocyst stage.

4 cells

8 cells

Morula 16-32 cells

Blastocyst

»Blastocyst Development

Steps towards implantation

During early blastocyst development, fluid starts to fill embryo. The embryo starts to expand like a balloon. To implant, the embryo must escape from the shell surrounding it. This is called "hatching".

Early Blastocyst

Late Blastocyst

Hatching Blastocyst

»Embryo Transfer

No place like home

• Embryos are transferred to the patient’s uterus on Day 3 or Day 5
• Most patients have Day 3 transfers
• Transferring later on Day 5 can help in embryo selection
• The deciding factor for a Day 3 vs Day 5 transfer is overall embryo number and cell division
• Extra non-transferred embryos of good quality are frozen

Day 3

Day 5

»Embryo Freezing

Planning for the future

• Extra embryos not used for the fresh transfer can be frozen between Day 3 and Day 6
• Only high quality embryos are frozen
• Vitrification a new advance in cryobiology is currently used for all embryo freezing
  
• Introduction of this new technique to our IVF lab has increased post - thaw survival and clinical pregnancy outcomes

»Egg Freezing

Storing fertility potential

• Patients may now freeze their unfertilized
  eggs for a future IVF cycle
• Egg freezing offers cancer patients an opportunity to bank oocytes before starting radiation or chemotherapy
• Ovarian tissue cryopreservation is also
  available for cancer patients

»Preimplantation Genetic Diagnosis (PGD)

Chromosomes tell all

• Screening embryos for genetic diseases
  is a powerful new tool to enhance opportunity for a healthy baby.
• CCF Fertility Center was the first program
  in northeast Ohio to offer PGD
• A single cell is removed on Day 3 at the 6-8 cell stage and its chromosomal status is assessed
• Only normal embryos are transferred
• Embryos can be screened for many diseases such as cystic fibrosis, muscular dystrophy, Down’s syndrome etc.

»Coculture for Implantation Failure

Helper cells to jumpstart the embryo

• Cleveland Clinic IVF program is one of the few centers offering co-culture to enhance embryo development
• Co-culture may benefit patients with previous failed IVF attempts or poor embryo quality
• Embryos are cultured in a unique chamber with uterine cells
• Growth factors from uterine cells may help struggling embryos to divide

»In Vitro Maturation

Growing eggs in a culture dish

• New frontier in assisted reproduction
• Immature oocytes recovered with minimal or no hormonal stimulation
• Immature oocytes cultured in the laboratory
• Benefits patients who don't respond well to conventional stimulations
• Reduce cost of IVF by minimizing use of medications