IVF Laboratory Technology - Week 1 Lecture Notes

Course Introduction and IVF Lab Workflow

Lecture Objectives

  • Understand course expectations.
  • Learn how the Skills Course is organized.
  • Learn key roles of the inter-professional team comprising an IVF facility.
  • Learn the basic workflow of the Embryology Lab.

Skills Development and Assessment

  • Goal: Provide students the opportunity to develop and assess skills needed and certify skill level.
  • Objectives: By the end of the course, students will develop basic skills through online and hands-on training at introductory, intermediate, or advanced levels, depending on the entering skill level.
  • Skill level will be assessed and certified.

IVF Skills Course

  • The online component supports the face-to-face session in July.
  • Broken down into techniques used in the Embryology and Andrology laboratories:
    • Semen Analysis
    • Sperm Prep
    • Embryo 101: microscopy, manipulation of oocytes and embryos
    • Embryo culture
    • Media and Media QC (pH, Temperature, osmolality)
    • Oocyte and Embryo Morphology
    • Time-lapse imaging
    • ICSI (Intracytoplasmic Sperm Injection)
    • Assisted Hatching
    • Embryo Biopsy
    • Vitrification of Oocytes and Embryos

Course Structure and Assessments

  • Each week in the online course introduces a technique or set of techniques using:
    • Lecture
    • Reading
    • In some cases, a discussion board.
  • Some weeks have self-assessment quizzes that are not graded.
  • Exams:
    • Three exams:
      • Week 5 (end of Module I)
      • Week 9 (just before the residential program, over Module II)
      • Week 11 (end of the residential, mock proficiency exam)
    • The final exam is cumulative (Module I, II, and all residential week material) and is given during week 12.

Required Textbook

  • David K. Gardner, Ariel Weissman, Colin M. Howles, and Zeev Shoham, Textbook of Assisted Reproductive Techniques Volume 1: Laboratory Perspectives, 5th edition, Informa, 2018, ISBN 13: ISBN 9781498740142
  • A PDF version is posted on the course site.
  • EVMS RCS Lab Manual Sections Posted In The Course Site.
  • Supplemental Readings: Selected papers as PDFs.
  • Supplemental Materials: Links to online materials may be posted.

Discussion Boards

  • Two online modules with deadlines:
    • Primary post by Saturday the discussion starts.
    • Secondary posts by Monday of the week the discussion starts, with the exception of the first discussion due on May 16th (Sunday).
  • Discussion Board Topics:
    • Course Expectations and Syllabus Review
    • Culture Discussion
    • Time Lapse Imaging
    • Embryos
    • Genetics

Exams Details

  • Three timed Exam Soft / Examplyfy Exams
  • Module I and Module II = 45 points each
  • Final Exam after the residential course = 50 points
  • Formats: Multiple choice, true/false, short answer, essay.

Skills Development and Assessment Details

  • Objectives:
    • Develop or enhance existing skills.
    • Certify skills to a mastered level (Expert, Intermediate, and Novice).
  • Each lab is worth 20 points across these categories:
    • Student knowledge of the technique
    • Demonstrated ability to learn and perform the task
    • Certification level as observed during the residential program

The IVF Team

  • Physician
  • Nurse Coordinator
  • Nurses
  • Andrology Lab
  • Embryology Lab
  • Endocrine Lab
  • Pre-Genetic Diagnosis
  • Research
  • Secretaries and Support Staff

Diagnosis and Treatment

  • Physicians diagnose infertility.
  • Involving laboratory personnel:
    • Andrology Lab: analysis of semen
    • Endocrinology: test hormone baseline levels to determine potential contribution
    • Embryology Lab: gamete isolation, in vitro fertilization, embryo evaluation and monitoring
  • Physicians determine the treatment protocol based on patient history and other diagnostic procedures.

Typical Workflow in IVF Lab

  • Patient undergoes 8 to 12+ days of hormone treatment (Stimulation) to mature oocytes.
  • About 35 hours before oocyte collection, a finalizing hormone shot (Trigger) is given to complete oocyte maturation.
  • Laboratory activity includes preparations for receiving oocytes the next morning:
    • Media preparation and overnight equilibration in the incubator.
  • Final Oocyte Maturation and HCG administration
  • Embryo Transfer
  • Progesterone supplementation
  • Pregnancy test

Day 0 - Laboratory Workflow

  • Day 0 - Transvaginal oocyte retrieval
    • Usually early in the morning, the patient undergoes the procedure to remove oocytes from her ovaries.
    • A transvaginal probe and needle, guided under ultrasound imaging, is used.
    • The needle aspirates the fluid along with the oocyte from the blister-like follicle.
    • The fluid is sent to the lab, where it is scanned to locate the cumulus-oocyte complex.

Types of Human Oocytes Recovered at Aspiration

  • Immature Complex - GV (Germinal Vesicle)
  • Mature Complex - PB (Polar Body)
  • Intermediate Egg
  • Atretic Egg

Day 0 - Sperm Collection

  • Count (number) & Motility (movement)

Day 0 - Insemination

  • Co-incubation of Sperm and Egg
  • In Vitro Fertilization (IVF) or ICSI (Intracytoplasmic Sperm Injection)

Oocyte Cryopreservation: Vitrification

  • If oocytes are not inseminated on Day 0, they can be preserved for future use via cryopreservation.
  • Vitrification: A rapid freezing technique that turns embryos into a glass-like state without ice crystal formation, preserving their viability.
  • Cryopreserved oocytes can be safely stored under liquid nitrogen in storage containers (dewars) for decades.

Day 1 – Assessment of Fertilization

  • Early the next morning (Day 1), the oocytes are assessed for fertilization.
  • Fertilized eggs are placed into culture.
  • ~16-19 hours post insemination: 2PN (2 pronuclei indicate fertilization)

Development of Human Embryos

  • Embryo Culture: 1-6 days
    • Day 1: 1-cell
    • Day 2: 2-4 cells
    • Day 3: 8-cell
    • Day 5 & 6: Blastocyst hatching

Embryo Development Assessment

  • Embryologists periodically assess embryo development by observing and recording the embryo’s morphology.
    • Number of blastomeres, their symmetry, and the degree of cytoplasmic fragmentation.
  • Embryos are judged by how “normal” their morphology and cleavage rates appear.
  • Balance between removing the embryo from the incubator environment and recording their development.
  • Time-lapse camera systems record embryo development within the incubator, eliminating the need to remove the embryo for observations.

Embryo Transfer

  • Transfer (could be Day 1-6)
    • Embryos are placed back into the patient.
    • GIFT: Egg & sperm into fallopian tube
    • ZIFT: Fertilized Egg into fallopian tube (TET)
    • IVF: Embryo into uterus
      • Day 2: 2-4 cells
      • Day 3: 8 cells
      • Day 5-6: Blastocyst

Assisted Hatching (AH) & Embryo Biopsy

  • Some programs prepare embryos for transfer by creating a hole in the zona pellucida – Assisted Hatching (AH).
  • AH is thought to aid the embryo in escaping from the zona pellucida, facilitating implantation in the uterine wall.
  • AH can also be performed to allow the embryologist to withdraw a cell or group of cells from the embryo for genetic testing – a procedure known as embryo biopsy.

Embryo Cryopreservation

  • Supernumerary embryos (those that meet usable quality criteria and are not intended for fresh transfer) can be safely stored for future use.
  • Embryos at any stage from zygote to blastocyst can be cryopreserved and safely stored in dewars for decades.
  • Vitrification Media:
    • Alcohols (dimethyl sulfoxide [DMSO], ethylene glycol, or propylene glycol) are the primary cryoprotectants.
    • Sugars (sucrose or trehalose) serve as osmoprotectants to help dehydrate the cells and protect membranes during freezing.