Embryo Biopsy - The Finer Points of Trophectoderm Biopsy - Suzanne Cawood CRGH ESHRE 2012 - RI presentation

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Information about Embryo Biopsy - The Finer Points of Trophectoderm Biopsy - Suzanne...
Health & Medicine

Published on December 20, 2013

Author: RI_UK

Source: slideshare.net

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The Finer points of Trophectoderm Biopsy Embryology, Suzanne Cawood Embryologist at CRGH - with lead Alpesh Doshi

The finer points of trophectoderm biopsy Suzanne Cawood The Centre of Reproductive and Genetic Health, London. UK

Preparing for blastocyst biopsy  Excellent blastocyst culture system - KPIs (Blastocyst formation rate, utilisation rate, %d5 vs. d6 formation rate  Group/individual culture until d5. If group culture, separate blasts morning of d5/6 into individual culture  Zona drilling d3 – ensure hole size optimum (too large, risk of premature cell loss; too small, risk of ‘pinching’ of trophectoderm cells) Biopsy in HEPES media. Make dishes 1 hour prior to procedure warmed at 37°C (non-gassed)  Ensure microscope stage heated to temperature which ensures droplet temperature is at 37 °C  Ensure optimum service is provided. Biopsy at d5AM/PM + d6 AM/PM is possible.

When and what to biopsy? WHEN?  Take into account how long the diagnosis will take.  Vitryfying blastocysts post biopsy? Or aiming for fresh embryo transfer? WHAT?  Hatching or Hatched blastocysts. Hatching more ideal  If herniating – trophectoderm cells will have to be ‘sucked’ from zona – more risk of lysis and damage to the embryo  ICM position

The procedure Rapid technique Minimise trauma/damage to both embryo and cells biopsied Use blastocyst holding pipette Common difficulties: Positioning of embryo/pipettes (Hatched blastocysts, ICM hatching out) Do not take too many cells! Remember image is 3D Control at all times – both holding, suction and laser. Utilise foot pedal if possible. Minimise number of laser shots – maximise shot value. Laser shots between the junction of trophectoderm cells Post biopsy, cells to be washed (PBS-PBA) and tubed (hand-pipetted) into empty eppendorf tubes

Technique of excising cells  Laser shot and tear cells  Fewer laser shots and ‘brushing’ the cells off using the holding pipette. - Difficulty: Cells are sticky! - more likely to leave cells on holding/aspiration pipette (potential contamination so ensure pipettes changed if necessary) - this technique has to be used for completely hatched blastocysts

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