ART- Oocyte Retrieval

lecture 4

How is oocyte retrieval preparation done based on cycle monitoring information in an IVF lab?

• hormone levels, follicular size and number estimate number of oocytes to be retrieved

• physician orders direct planning and preparation regarding procedures and techniques

  • type of insemination (ISCI, conventional)

  • fresh vs frozen transfer

  • assisted hatching

  • PGT

What four main processes must be dine for worksheet and dish prep?

1. patient ID labelling

2. embryology treatment verification

3. prepared worksheets of embryo scheduling

4. equilibration of media and dishes

What steps are included in labelling patient dishes?

Labels on top and bottom of dish and at beats two unique identifiers: Name, DOB, chart #

How are patient dishes organized and verified within the IVF lab?

Dishes have preassigned location in incubator, with electronic tracking verification

How are treatment orders and dish/media prepped prior to retrieval?

Treatment order helps to organize what dishes/media needs to be prepared and how much, must be done prior to retrieval as media takes several hours to equilibrate

How might media be mimicked to match follicular fluid?

Temp, pH, Osmolarity

• Hyaluronidase for ICSI to denude cumulus cells, bc found in vivo to help expand cumulus cells for fertilization

What is a “time-out” when it comes to actual oocyte retrieval?

• Meeting between lab and physician to verify patient, orders, treatment

• verify tube warmers (37 ± 0.5c) and flush media pH

• suction pressure (1000-140 mmHg)

How large are COCs?

5mm

What media are COCs retrieved in?

Flush media (contains insemination media, stable pH (HEPEs/MOBs)), and temp)

How are COCs removed from flush media?

Large pipettes careful not to damage oocyte

COCs are removed from flush media and transferred to ______ media?

Culture media

How are COCs assessed?

Based on expansion, appearance of cumulus cells, and size

What does an immature vs atretic COC look like?

immature- small, dense cumulus cells, no germinal vesicle, no PB

atretic- small, dark, few cumulus cells

What does mature COCs look like?

expanded cumulus

distinct ZP

clear ooplasm

large

How large is an oocyte?

0.1mm

What is the process of denuding/ stripping COCs?

exposure to hyaluronidase to loosen cumulus cells and pulling complex in and out of pipette to remove cells

How long after retrieval is fertilization done?

1-4 hours for conventional or ICSI

Trouble shooting techniques for a failed oocyte retrieval?

1. Double check monitoring info… maybe no large ones anyway

2. Follicular cells in fluid?

3. was trigger taken properly?

4. Consider options at ICSI and fertilization next day