6/24/2019 SPERM, OOCYTES AND EMBRYOS SPERM, OOCYTES AND EMBRYOS SPERM, OOCYTES AND EMBRYOS SPERM, OOCYTES AND EMBRYOS IN THE IVF LAB IN THE IVF LAB IN THE IVF LAB IN THE IVF LAB JILL MELLON, BSc.(Hons), MSc. Lab Director PCRM Burnaby JASON AU, BSc., MSc. Lab Supervisor PCRM Burnaby 1 www.pacificfertility.ca We will discuss: We will discuss: We will discuss: We will discuss: 1 Sperm Basics of Semen Analysis - what it tells us • Most suitable treatment - guidelines • Oocytes 2 Ovarian stimulation to produce more eggs • Egg numbers and quality • IVF and ICSI 3 IVF • ICSI • Embryo Development • Biopsy of embryos for Genetic Testing 4 Purpose • Procedure • 2 www.pacificfertility.ca 1
6/24/2019 INTRODUCTION INTRODUCTION INTRODUCTION INTRODUCTION All staff have at least a BSc. in a Science subject and some of us have a Masters in Clinical Embryology Combined experience of more than 60 years (not all me!) Training is on site and extensive: normally ONE year for Andrologist and TWO years for Embryologist Known within the Clinic for our attention to detail which borders on OCD Compassionate and caring people with an unhealthy interest in our results! 3 www.pacificfertility.ca 1. Sperm 1. Sperm 1. Sperm 1. Sperm Basic Semen Analysis • Abstinence (2-5 days) • Time to when examination is performed (30 mins) • Volume = semen volume (>1.5mL) • Concentration = how many millions per ml (>15) • Sperm Motility = good, fair, poor (≥40%) • Morphology = normal or abnormal (≥4%) Semen analysis is a good non-invasive test to assist the Physician in directing optimal treatment 4 www.pacificfertility.ca 2
6/24/2019 WHO Guideline WHO Guideline WHO Guideline WHO Guideline “Data characterizing the semen quality of fertile men, whose partners had a time to pregnancy of 12 months or less, provided the reference ranges for this manual. Raw data from between about 400 and 1900 semen samples, from recent fathers in eight countries on three continents, were used to generate the reference ranges.” 5 www.pacificfertility.ca Sperm Morphology Sperm Morphology Sperm Morphology Sperm Morphology WHO Laboratory Manual for the Examination and Processing of Human Semen. 2010. 6 www.pacificfertility.ca 3
6/24/2019 Sperm Sperm Sperm Sperm NOTE: Sperm production takes 65-80 days illness may affect the sample – therefore repeat if necessary Always consider male and female factors (such as duration of infertility, patient age and partner age, previous pregnancy) first choice of treatment must include consideration of all factors REI recommendation is for what will give highest chance of success in our IVF Lab, and does not always mean other treatments cannot work Recommended treatments: intrauterine insemination, Standard IVF insemination, and intracytoplasmic sperm injection. 7 www.pacificfertility.ca Sperm Washing for Insemination Sperm Washing for Insemination Sperm Washing for Insemination Sperm Washing for Insemination YouTube “PCRM: The IVF Laboratory” https://www.youtube.com/watch?v=XFE-kd0nJRM 8 www.pacificfertility.ca 4
6/24/2019 Sperm Washing for Insemination Sperm Washing for Insemination Sperm Washing for Insemination Sperm Washing for Insemination Pre-wash Post-wash 9 www.pacificfertility.ca Pregnancy Rates by Treatment Type Pregnancy Rates by Treatment Type Pregnancy Rates by Treatment Type Pregnancy Rates by Treatment Type 70% 60% 57% 52% 50% Clinical Pregnancy Rate 43% 38% 40% IUI-H 30% 25% IUI-D 21% IVF/ICSI 20% 15% 14% 13% 12% 8% 8% 10% 5% 3% 0% 0% <35 35-37 38-40 41-42 >43 Age Groups 10 www.pacificfertility.ca 5
6/24/2019 Sperm Freezing Sperm Freezing Sperm Freezing Sperm Freezing Sperm can be frozen indefinitely and stored in liquid nitrogen, with no effect on its quality. Semen has a cryoprotectant solution added before being frozen in “straws” using a controlled rate freezer. Sperm freezing is done for many reasons : As back up for a treatment cycle, or when the patient will be unavailable during treatment or has problems producing a sample For cancer patients, sperm can be frozen pre-treatment to preserve fertility. This includes adolescent patients Transgender patients can freeze sperm before their gender re-assignment 11 www.pacificfertility.ca 2. Oocytes (Eggs) 2. Oocytes (Eggs) 2. Oocytes (Eggs) 2. Oocytes (Eggs) MI GV MII Hill, M.A. (2019, May 19) Embryology Female gametogenesis.jpg . Retrieved from https://embryology.med.unsw.edu.au/embryology/index.php/File:Female_gametogenesis.jpg 12 www.pacificfertility.ca 6
6/24/2019 Egg Retrieval Egg Retrieval Egg Retrieval Egg Retrieval 13 www.pacificfertility.ca Egg Masses Egg Masses Egg Masses Egg Masses – – Cumulus Oocyte Complex – – Cumulus Oocyte Complex Cumulus Oocyte Complex Cumulus Oocyte Complex Average number of eggs retrieved in <35yo = 16 Average number of eggs retrieved in >40yo = 9 14 www.pacificfertility.ca 7
6/24/2019 Oocytes Maturity Oocytes Maturity Oocytes Maturity Oocytes Maturity Zhang P, Zucchelli M, Bruce S, et al. Transcriptome profiling of human pre-implantation development. PLoS ONE. 2009;4(11):e7844. 15 www.pacificfertility.ca Oocytes Quality Oocytes Quality Oocytes Quality Oocytes Quality Poor quality oocyte Good quality oocyte 16 www.pacificfertility.ca 8
6/24/2019 We will discuss: We will discuss: We will discuss: We will discuss: Sperm 1 Basics of Semen Analysis - what it tells us • Most suitable treatment - guidelines • Oocytes 2 Ovarian stimulation to produce more eggs • Egg numbers and quality • IVF and ICSI 3 IVF • ICSI • Embryo Development • 4 Biopsy of embryos for Genetic Testing Purpose • Procedure • 17 www.pacificfertility.ca IVF A calculated number of highly motile sperm is added to a dish containing the eggs. This is incubated overnight and checked the following morning for signs of fertilization Used where semen analysis is in the normal range Considered to be more “natural” and less invasive than ICSI 18 www.pacificfertility.ca 9
6/24/2019 ICSI (Intracytoplasmic Sperm Injection) ICSI (Intracytoplasmic Sperm Injection) ICSI (Intracytoplasmic Sperm Injection) ICSI (Intracytoplasmic Sperm Injection) A single sperm is injected Reasons where the preferred into a single mature egg. treatment may be ICSI Low concentration of sperm No normal sperm Extracted sperm Poor sperm motility To optimize (not guarantee) fertilization – for example for biopsy and genetic testing 19 www.pacificfertility.ca IVF/ICSI IVF/ICSI – IVF/ICSI IVF/ICSI – – EMBRYO DEVELOPMENT – EMBRYO DEVELOPMENT EMBRYO DEVELOPMENT EMBRYO DEVELOPMENT 20 www.pacificfertility.ca 10
6/24/2019 IVF/ICSI IVF/ICSI - - Embryo Development: Blastocysts Embryo Development: Blastocysts IVF/ICSI IVF/ICSI - - Embryo Development: Blastocysts Embryo Development: Blastocysts YouTube “Reproductive System, Part 4 - Pregnancy & Development: Crash Course A&P #43” https://www.youtube.com/watch?v=BtsSbZ85yiQ 21 www.pacificfertility.ca IVF/ICSI – IVF/ICSI IVF/ICSI IVF/ICSI – – TRANSFER AND FREEZE – TRANSFER AND FREEZE TRANSFER AND FREEZE TRANSFER AND FREEZE Only a proportion of these will become blastocysts We very rarely transfer more than 2 embryos, and recommend transfer of only 1 in patients up to age 37 The embryo with best “morphology” is transferred and others which make good quality blastocysts are vitrified Damage or loss are rare when they are warmed, and pregnancy rates are the same 22 www.pacificfertility.ca 11
6/24/2019 IVF/ICSI FREEZE (VITRIFICATION) IVF/ICSI FREEZE (VITRIFICATION) IVF/ICSI FREEZE (VITRIFICATION) IVF/ICSI FREEZE (VITRIFICATION) Freezing (Vitrification) The process takes 30-40 minutes for up to 4 embryos Embryos are stored in straws, within small tanks filled with liquid nitrogen 23 www.pacificfertility.ca IVF/ICSI FREEZE AND THAW IVF/ICSI FREEZE AND THAW IVF/ICSI FREEZE AND THAW IVF/ICSI FREEZE AND THAW Thawing (Warming) The process takes around 30 minutes for each embryo and then the embryo is given a few hours to recover before being transferred. Immediately post-thaw 3 hours post-thaw 24 www.pacificfertility.ca 12
6/24/2019 IVF/ICSI – IVF/ICSI – PREGNANCY RATES FRESH VS. FROZEN PREGNANCY RATES FRESH VS. FROZEN IVF/ICSI IVF/ICSI – – PREGNANCY RATES FRESH VS. FROZEN PREGNANCY RATES FRESH VS. FROZEN Fresh V. Frozen Transfer - CPR 2018 60 50 40 30 20 10 0 <35 years 35-37 years 38-40 years fresh ET frozen ET 25 www.pacificfertility.ca 4. Embryo Biopsy for preimplantation genetics testing 4. Embryo Biopsy for preimplantation genetics testing 4. Embryo Biopsy for preimplantation genetics testing 4. Embryo Biopsy for preimplantation genetics testing Why PGT? • Previous pregnancy losses • Advanced maternal age • Familial genetic condition • Multiple failed transfers from IVF cycles Reduce TTP Save storage of abnormal embryos Family planning Surrogate community Frozen donor eggs 26 www.pacificfertility.ca 13
6/24/2019 Biopsy Biopsy Biopsy Biopsy 27 www.pacificfertility.ca Thank you to our amazing team! Thank you to our amazing team! Thank you to our amazing team! Thank you to our amazing team! 28 www.pacificfertility.ca 14
6/24/2019 WE WELCOME YOUR WE WELCOME YOUR WE WELCOME YOUR WE WELCOME YOUR QUESTIONS! QUESTIONS! QUESTIONS! QUESTIONS! 15
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