Applicants will be expected to have a good knowledge of the following aspects of clinical embryology.
Basic Cell Biology
1.1 The cell
- Internal organisation
- Cell cycle control, checkpoints
- Mitosis and meiosis
- Reproductive cells: spermatozoa and oocytes
1.2 Cell-cell interaction
- Membrane receptors: function, type, regulation
- Signalling
- Junctions
1.3 Basic genetics of the cell
- DNA chromatin and chromosomes
- Concept of a gene
- Mutations
- Epigenetics
1.4 Basic gene regulation
- Translation
- Transcription
- Expression
- Imprinting
Genetics
2.1 Basic genetics principles
- Genotype and phenotype
- Basic Mendelian inheritance patterns
- Monogenic diseases
- Mutations, copy number variation (CNV), de novo mutations
- Chromosomal abnormalities: numerical, structural
- Interpretation of an inheritance / family tree / pedigree
2.2 Genetic analysis and diagnosis
- How and why is it performed
- Diagnostic methods: cytogenetics (e.g. karyotyping, FISH), molecular genetics (e.g. PCR, array CGH, NGS)
Developmental Biology
3.1 Embryonic stem cells
- Origins, definitions, characteristics
3.2 The fetal ovary
- Factors regulating development
- Primordial germ cells
- Cell migration
- Time scale (days / week)
3.3 The fetal testis
- Factors regulating development
- Primordial germ cells
- Cell migration
- Time scale (days / week)
3.4 Gamete interaction – until 1st cleavage
- Fertilisation
- Acrosome reaction
- Sperm- oocyte signaling
- Sperm decondensation
- Oocyte activation
- Meiosis II, pronuclei and spindle formation
3.5 Embryo development – from first cleavage to implantation
- Metabolism, cell positions, embryonic axis
- Kinetics, timing, regulation
- Apoptosis
3.6 Implantation
- Hatching, adhesion, invasion, endometrium
3.7 Post-implantation embryology
- Gastrulation
- Organogenesis
- Sex differentiation
3.8 Early pregnancy
- hCG production, biochemical and ongoing pregnancy
- Implantation, ultrasound (sacs, heartbeat)
- Extra uterine pregnancies
- Spontaneous abortions
- Embryo factors vs. uterine factors in implantation/implantation failure
Female Reproduction
4.1 Anatomy and function of the female reproductive system
- Role of accessory systems
- Function of the organs
4.2 Oogenesis
- Regulating factors
- Hypothalamus, pituitary, gonad axis
- Endocrine regulation
- Theca & granulosa cells
- Maturation biochemistry and metabolism of the oocyte
- Oocyte morphology/structure
- Function of each structure
4.3 The oocyte: markers of competence
- Nuclear maturity
- Cytoplasm
- Polar bodies
- Zona pellucida
- Cumulus cells
4.4 Clinical workout
- Evaluation of (in)fertility: aetiology, medical/physical aspects, genetic diagnosis, hormonal evaluation, treatment options, etc.
- Definitions, primary infertility, secondary infertility
- Serological screening for patients and/or donors
- Effects of treatment
- Oocyte donation
4.5 Ovarian hyperstimulation
- Basic principles
- Types of medication
- Stimulation regimes (types, rationales)
- Complications of treatment
Male Reproduction
5.1 Anatomy and function of the male reproductive system
- Role of accessory systems
- Function of the organs
5.2 Spermatogenesis
- Regulating factors
- Hypothalamus, pituitary, gonad axis
- Endocrine regulation
- Leydig & Sertoli cells
- Differentiation and maturation
- Biochemistry and metabolism of the sperm cell
- Sperm morphology/structure
- Function of each structure
5.3 Diagnosis of male infertility
- Semen analysis (functional analysis, microscopic analysis)
- WHO & ESHRE guidelines
- CASA systems
5.4 Clinical workout
- Evaluation of (in)fertility: aetiology, medical aspects, genetic diagnosis, hormonal evaluation, physical aspects, treatment options, etc.
- Definitions, primary/secondary infertility
- Serological screening for patients and/or donors
- Sperm donation
MAR Laboratory Procedures
6.1 Strategies for choosing fertilization procedures
- IUI, IVF or ICSI, criteria
- IVM
- PESA, TESA, TESE
- Donor sperm in relation to serological tests (different handling and storage)
6.2 The sperm sample: preparation methods
- Gradient centrifugation, swim-up, swim-out, etc
- Advanced diagnostic tests (evaluation of DNA damage, chromatin condensation, etc)
- When to use what, why, differences
6.3 MAR techniques
- Practicalities for IUI, IVF and ICSI (timing, preparation procedures, materials, etc)
- Pick-up, oocyte handling
- IVF insemination
- Denudation prior to ICSI
- ICSI procedure
6.4 Embryo scoring, Day 1 – 6
- PN scoring
- Morphology criteria
- Kinetics, genetics, physiology (e.g. amino acids, oxygen metabolism)
- Time-lapse
- Destination of embryos: selection criteria for transfer, cryopreservation, biopsy, etc
6.5 Embryo transfer
- Identity check
- Selection and criteria for number of embryos to be transferred
- Catheter loading and transfer procedure
- Assisted hatching: pros-cons, evidence
6.6 PGT
- Timing of biopsy
- Tubing techniques
- Zona opening (pros and cons)
- Different biopsy types, number of cells to evaluate
- Techniques used for evaluation of the cells biopsied
- Results and recommendations upon transferring embryos after PGT
6.7 Culture conditions
- Media
- Type of culture systems
- Requirements for consumables
- Physiochemical parameters (temperature, pH, osmolality)
- Stage specific requirements
6.8 Non-routine methods:
- Examples of non-routine methods, e.g. in vitro maturation, polar body evaluation, assisted hatching, etc
Cryopreservation
7.1 Principles of cryopreservation
- Basic cryobiology
- Cryoprotectants, additives
- Slow freezing
- Vitrification
- Advantages/disadvantages with different methods
- Fertility preservation in oncology cases
7.2 Cryopreservation of sperm
7.3 Cryopreservation of oocytes
7.4 Cryopreservation of embryos
7.5 Cryopreservation of ovarian tissue
7.6 Cryopreservation of testicular tissue
7.7 Equipment
- Machines
- Straws/ampoules
- Contamination risk from storage medium
- Minimal safety requirements
- Security
7.8 The cryopreservation-thawed/warmed embryo treatment cycle
- Monitoring and timing of the thawing/warming cycle
- Controlled and natural cycles
Laboratory and Quality Management
8.1 Patient data
- Identity check
- Confidentiality
- Keeping records
- Safety, storage of data
- Single European Code
8.2 Type and treatment choices
- Surgical
- Hormone stimulation
- Insemination IVF / ICSI
- Use of fresh or frozen gametes
- Gamete and embryo donation
8.3 Treatment outcome
- The health of the children
- Risk factors
- Maternal factors
- Paternal factors
- Multiple pregnancies
- Chromosomal factors
- Malformations
- Imprinting
8.4 Quality assurance
- Identification procedures
- Laboratory and clinical KPIs
- Standard operating procedures
- Traceability of samples/material
- Validation procedures
- Risk analysis
- Logbooks
- Introducing new methods
8.5 Equipment and facilities
- Calibrations
- Validation, monitoring, logbooks, maintenance and control
- Microscopes
- Principals of optical system, calibrations, maintenance and control
- Technical requirements in a MAR lab facility
8.6 Statistical analysis
- Sample size evaluation
- Study design
- Statistical variance
- Interpretation of results
8.7 Legislation
- ESHRE guidelines
- Ethical considerations
- Code of practice
- European rules and regulations related to MAR practice
- EU Tissue and Cells Directives (EUTCDs)
8.8 Reducing risks/ dealing adverse events/ troubleshooting
- Contaminated samples
- Processing and storage of contaminated samples with contagious agents
- Personal protective equipment (PPE)
- Hygiene and disinfectants used at MAR lab
- Protective measures (alarm, sensors, etc)
- Actions upon injury
- Risk of mix-up of gametes, loss or damage during handling
- Transfer of wrong embryos
- Breakdown of equipment, back-up strategies
