Mastering Clinical Embryology (Mar 22, 2024)_(1032216743)_(CRC Press)

Cover
Half Title
Title
Copyright
Contents
List of Videos
Foreword
List of Contributors
1 Laboratory Design
	1.1 Planning the IVF Laboratory
	1.2 Equipment
	1.3 Cryolaboratory
	1.4 Future-Proofing
	1.5 Location of the Laboratory
	1.6 Materials
	1.7 Other Considerations
	1.8 Final Word
	References
2 IVF Cell Culture: VOCs and Air Quality
	2.1 What Are VOCs?
	2.2 Methods of Measuring VOCs
		2.2.1 PID Sensors
		2.2.2 Canister-Based Sampling
	2.3 VOCs and IVF
	2.4 Other Airborne Contaminants
	2.5 VOC Remediation Methods
		2.5.1 Potassium Permanganate-Impregnated Activated Carbon
		2.5.2 Photocatalytic Oxidation (PCO)
		2.5.3 IVF-Purposed Air Purification: LifeAire System
	2.6 Summary
	References
3 Laboratory Equipment and Consumables
	3.1 Equipment
		3.1.1 Laminar Flow Hoods
		3.1.2 Microscopes
		3.1.3 Incubators
		3.1.4 Cryostorage
		3.1.5 Centrifuges
		3.1.6 Refrigerators and Freezers
		3.1.7 Other Supplementary Equipment
		3.1.8 Monitoring and Alarm Systems
		3.1.9 Quality Management Equipment
	3.2 Consumables
		3.2.1 Plasticware
		3.2.2 Culture Media
		3.2.3 Oil for Embryo Culture
		3.2.4 Flushing Media
		3.2.5 Cryopreservation Media and Consumables
		3.2.6 Liquid Nitrogen
		3.2.7 Micromanipulation Pipettes
		3.2.8 Gases
		3.2.9 Oocyte Aspiration Needles
		3.2.10 Embryo Transfer Catheters
	3.3 Stock Management
	3.4 Contingency Equipment
	3.5 SOPs, Instruction Manuals and Cleaning Regimes
	3.6 Electronic Witnessing
	3.7 Final Word
	References
4 Staffing in the IVF Laboratory
	4.1 Training to Work in an IVF Laboratory
	4.2 Staff Numbers and Skills
	4.3 IVF Laboratory Team Roles and Ratios
	4.4 Calculating the IVF Lab Staff Required Based on Workload and Other Factors
	4.5 Finally on Recruitment and Retention
	References
5 Evidence-Based Practice in Reproductive Medicine
	5.1 Introduction
	5.2 What Is Evidence-Based Practice?
	5.3 Step 1: Translation of Uncertainty into an Answerable Question (Ask)
	5.4 Step 2: Systematic Retrieval of Best Evidence Available (Acquire)
	5.5 Step 3: Critical Appraisal of Evidence for Validity, Clinical Relevance, and Applicability (Appraise)
	5.6 Step 4: Application of the Results in Practice (Apply)
	5.7 Step 5: Evaluation of Performance (Audit)
	5.8 Conclusion
	References
6 Quality Control and Management
	6.1 Introduction
		6.1.1 Quality Control and Quality Assurance
		6.1.2 Quality Management and Regulation
	6.2 The Quality Manual
	6.3 The Quality Policy
	6.4 Quality Objectives
	6.5 Establishing the QMS
		6.5.1 Document Management
		6.5.2 Standard Operating Procedures
		6.5.3 Audit Documentation
	6.6 Implementing and Maintaining the QMS
		6.6.1 User Feedback
		6.6.2 Audit
		6.6.3 Key Quality and Performance Indicators
		6.6.4 Change Control
		6.6.5 Continual Improvement
	6.7 Why Does Quality Management Matter?
	6.8 Appendix: Definitions
	References
7 Analysis, Reporting and Presentation of Findings
	7.1 Analysis
		7.1.1 The Null Hypothesis
		7.1.2 Testing the Hypothesis
		7.1.3 Sample Sizes
		7.1.4 Retrospective Analyses
		7.1.5 Statistical Analyses
		7.1.6 Ethical Approval
	7.2 Reporting
	7.3 Presentation
		7.3.1 Writing an Abstract
		7.3.2 Presenting a Poster
		7.3.3 Presenting Orally
		7.3.4 Writing a Manuscript
	7.4 Conclusion
	References
8 The Human Fertilisation and Embryology Authority (HFEA)
	8.1 Licensing of ART Providers
	8.2 The HFEA Code of Practice
	8.3 Consent: Enshrining the Rights of Egg and Sperm Providers
	8.4 Consent for Treatment
	8.5 Consent to Storage
	8.6 Becoming the Legal Parent of a Child
	8.7 Consent to Disclose of Identifying Information: Enshrining Privacy and Confidentiality
	8.8 Counselling and Information: The Right to Be Fully Informed
	8.9 Welfare of the Child
	8.10 Treatment with Donated Eggs Sperm and Embryos
	8.11 Embryo Testing
	8.12 Training and Research
	8.13 Summary
	References
9 Anatomy and Physiology of Mammalian Reproductive System
	9.1 Anatomy of Female Reproductive System
		9.1.1 External Genitalia (Vulva)
		9.1.2 Uterus and Cervix
		9.1.3 Fallopian Tubes (Uterine Tubes)
		9.1.4 Ovaries
		9.1.5 Vagina
	9.2 Anatomy of Male Reproductive System
		9.2.1 Testes
		9.2.2 Epididymis
		9.2.3 Accessory Glands
		9.2.4 Penis
	9.3 Physiology of the Female Reproductive System
		9.3.1 Regulation of Menstrual Cycle
	9.4 Physiology of the Male Reproductive System
		9.4.1 Spermatogenesis
		9.4.2 Spermiogenesis
		9.4.3 Maturation
		9.4.4 Hormonal Regulation of Spermatogenesis and Steroidogenesis
		9.4.5 Erection and Ejaculation
	References
10 Epigenetics and ARTs
	10.1 Introduction
	10.2 Epigenetics and Development
	10.3 Epigenetics and ART
	10.4 Conclusion
	References
11 Folliculogenesis and Oogenesis
	11.1 Introduction
	11.2 Folliculogenesis
		11.2.1 Primordial Follicle Activation
		11.2.2 Preantral Follicular Development: Primary to Secondary Follicle
		11.2.3 Antral Follicular Development
		11.2.4 Antral Follicular Recruitment
		11.2.5 Selection of the Dominant Follicle
		11.2.6 Follicular Dominance
		11.2.7 Intra-Ovarian Factors
	11.3 Oogenesis
	11.4 Conclusions
	References
12 Spermatogenesis, Spermiation and the Human Sperm
	12.1 Introduction
	12.2 Physiology of the Testis and Seminiferous Tubule
	12.3 Spermatogenesis and Spermiogenesis
	12.4 The Leydig Cell
	12.5 The Sertoli Cell
	12.6 The Blood-Testis Barrier (BTB)
	12.7 The Structure of the Mature Sperm
	12.8 Post-Ejaculation Changes
	12.9 Conclusions
		12.9.1 Funding
	References
13 Fertilisation and Pre-Implantation Embryo Development
	13.1 Scientific Theory Underpinning Practice
		13.1.1 Signs of Fertilisation
		13.1.2 Pronuclei Appearance
		13.1.3 Laboratory Considerations
		13.1.4 Triploid Embryos
	13.2 Diandric Embryos (1 Maternal, 2 Paternal PN)
	13.3 Digynic Embryos (2 Maternal and 1 Paternal PN)
	13.4 Implications
		13.4.1 Artificial Oocyte Activation (AOA)
		13.4.2 Embryo Development
		13.4.3 Pronuclei Disappearance and Mitotic Cleavage Divisions
		13.4.4 Morula and Blastocyst
		13.4.5 Aneuploidy and Morphokinetic Abnormalities
		13.4.6 Embryo Grading
		13.4.7 Blastocyst Grading
	References
14 The Endometrium, Implantation and Placental Development
	14.1 The Endometrium
		14.1.1 Physiology of the Endometrium through the Menstrual Cycle
		14.1.2 Endometrial Receptivity
	14.2 Embryo Implantation
	14.3 Placental Development
	14.4 The Endometrium in Assisted Reproduction
	14.5 Conclusion
	References
15 Fetal Development and Sex Differentiation
	15.1 Introduction
	15.2 Sexual Determination
	15.3 Gonadal Differentiation: Testis
	15.4 Gonadal Differentiation: Ovary
	15.5 Hypothalamic Sex Differentiation: GnRH Release
	15.6 Conclusion
	References
16 Recurrent Implantation Failure and Miscarriage
	16.1 Definition
		16.1.1 Recurrent Implantation Failure (RIF)
		16.1.2 Miscarriage
	16.2 Prevalence Risk
	16.3 Aetiology and Management
		16.3.1 Embryonic Factors
		16.3.2 Non-Embryonic Factors
	16.4 Conclusion
	References
17 Infertility: Definition, Referral, and Initial Consultation
	17.1 Referral to a Fertility Specialist
	17.2 History Taking
	17.3 Baseline Investigations
		17.3.1 Testing for Ovarian Reserve
		17.3.2 Basal FSH
		17.3.3 AMH
		17.3.4 AFC
		17.3.5 Investigating Ovulation
		17.3.6 Tubal Patency Studies
		17.3.7 HSG
		17.3.8 HyCoSy
		17.3.9 Laparoscopy and Dye
		17.3.10 Semen Analysis
	References
18 Ovarian Stimulation Protocols
	18.1 Introduction
	18.2 Physiology of Ovarian Stimulation
	18.3 Oral Agents for Ovulation Induction
		18.3.1 Clomifene Citrate
		18.3.2 Letrozole
		18.3.3 Metformin
	18.4 Gonadotrophins for Ovarian Stimulation
		18.4.1 Urinary Gonadotrophins
		18.4.2 Recombinant Gonadotrophins
		18.4.3 Effectiveness and Safety of Exogenous Gonadotrophins
	18.5 Gonadotrophin-Releasing Hormone and Analogue Drugs
		18.5.1 GnRH Agonists
		18.5.2 GnRH Antagonists
	18.6 Controlled Ovarian Stimulation Protocols
		18.6.1 Long GnRH Agonist Protocols
		18.6.2 Short GnRH Antagonist Protocols
		18.6.3 GnRH Agonist Flare Short Protocols
		18.6.4 GnRH Agonist Flare Ultrashort Protocols
		18.6.5 Relative Effectiveness and Safety of COS Protocols
	18.7 Conclusion and Future Perspectives
	References
19 Oocyte Retrieval and Evaluation
	19.1 Development of Modern-Day Techniques
	19.2 Procedural Timing
	19.3 Oocyte Retrieval
		19.3.1 Theatre Equipment
		19.3.2 Laboratory Equipment
		19.3.3 Ovarian Reserve and Yield
		19.3.4 Ovarian Cysts
	19.4 Oocyte Handling and Maintaining Gamete Viability
		19.4.1 Temperature
		19.4.2 pH
		19.4.3 Osmolality
	19.5 In Vitro Culture and Preparation for Insemination
	19.6 Oocyte Evaluation
		19.6.1 Cumulus-Oocyte Complex
	19.7 Oocyte Maturity
		19.7.1 Germinal Vesicle
		19.7.2 Metaphase II
		19.7.3 Metaphase I
	19.8 Oocyte Quality
		19.8.1 Shape
		19.8.2 Zona Pellucida
		19.8.3 Polar Body
		19.8.4 Dark Cytoplasm or Granularity
		19.8.5 Smooth Endoplasmic Reticulum Discs
		19.8.6 Vacuoles
		19.8.7 Perivitelline Space
		19.8.8 Membrane Behaviour at ICSI
		19.8.9 Aneuploidy
	19.9 Conclusion
	References
20 Semen Analysis, Preparation, and Selection
	20.1 Semen Analysis
		20.1.1 Critical Patient Information
	20.2 Macroscopic Evaluation
	20.3 Microscopic Evaluation
		20.3.1 Calculating the Concentration
		20.3.2 Suspected Azoospermia
		20.3.3 Assessing the Motility
		20.3.4 Categories of Sperm Motility
		20.3.5 Assessing the Morphology
	20.4 Sum of All Abnormalities/Number of Abnormal Sperm
		20.4.1 Round Cells
		20.4.2 Reporting and Interpreting the Results
	20.5 Preparation of Semen for Treatment
		20.5.1 Wash
		20.5.2 Swim Up
		20.5.3 Density Gradient
		20.5.4 The Future of Sperm Selection and Preparation for Treatment
	20.6 Conclusion
	References
21 Fertilisation Methods
	21.1 Introduction
	21.2 Intrauterine Insemination (IUI)
	21.3 In Vitro Fertilisation (IVF)
		21.3.1 Short Incubation Time
	21.4 Intra Cytoplasmic Sperm Injection
		21.4.1 History
		21.4.2 When to Use ICSI
		21.4.3 The Equipment Required for ICSI
		21.4.4 Stages in the ICSI Process
	21.5 Conclusion
	References
22 Embryo Culture Methods and Metabolic Requirements
	22.1 Metabolic Requirements
		22.1.1 Energy Source
		22.1.2 Pyruvate
		22.1.3 Lactate
		22.1.4 Glucose
		22.1.5 Amino Acids
		22.1.6 Other Micronutrients and Antioxidants
		22.1.7 Antibiotics
		22.1.8 Buffers
		22.1.9 Macromolecules
	22.2 Embryo Culture/History and Commercialisation
	22.3 Types of Media
		22.3.1 Single Step
		22.3.2 Sequential Media
		22.3.3 Suites of Media Tailored to a Specific Process
	22.4 Incubation
		22.4.1 Temperature
		22.4.2 CO2/O2 Gas Regulation and pH
		22.4.3 Oxygen Concentration
		22.4.4 Air Quality
		22.4.5 Humidity
	22.5 Media Storage
	22.6 Dish Preparation
		22.6.1 Dishes and the Microenvironment
	22.7 Developing Areas of Embryo Culture
	22.8 Conclusion
	References
23 Ethics and the Law in Embryology
	23.1 Introduction
	23.2 Embryology and the Law in the UK
	23.3 Why Is Fertility Medicine Different?
	23.4 Considering Possible People
	23.5 Ethical Approaches
		23.5.1 Consequentialism
		23.5.2 Comparisons with Adoption
		23.5.3 Rights-Based Systems of Ethics
		23.5.4 Fairness
	23.6 The Legal and Moral Status of the Human Embryo
	23.7 Ethical Considerations Around Embryonic Stem Cells
	23.8 Confidentiality and Infertility
	23.9 Ethical Dilemmas Arising from the Cryopreservation of Gametes
		23.9.1 Posthumous Reproduction and the Cryopreservation of Gametes
		23.9.2 Posthumous Children
		23.9.3 Controversies in ‘Social’ or ‘Elective’ Egg Freezing
	23.10 Donor Gametes: Ethical Issues for Donors, Recipients and Their Offspring
		23.10.1 Surrogacy
	23.11 ‘Designing Babies’: Balancing Autonomy with Welfare of the Child
	23.12 Conclusion
	References
24 The Patient Experience through an IVF Treatment Cycle
	24.1 How Do Patients Select a Fertility Clinic?
	24.2 Getting Started
	24.3 Modes of Communication with Patients
	24.4 Funding Treatment
	24.5 During IVF Treatment
	24.6 The Two-Week Wait
	24.7 Patient Support
		24.7.1 Fertility Support Events
		24.7.2 Buddy Systems
		24.7.3 Forums
		24.7.4 Support Counselling
	24.8 Concluding Thoughts on Creating Families
25 Non-Invasive Markers of Gamete and Embryo Quality
	25.1 Introduction
	25.2 Traditional and Timelapse Observations
	25.3 Use of Machine Learning and Artificial Intelligence
	25.4 Microscopic Advances
	25.5 Intracytoplasmic Morphologically Selected Sperm Injection (IMSI)
	25.6 Analysis of Spent Culture Media
	25.7 Non-Invasive PGT-A
	25.8 Summary
	References
26 Embryo Transfer Procedures and Decisions
	26.1 Timing of Embryo Transfer
		26.1.1 Blastocyst Transfer
		26.1.2 Cleavage Stage Embryo Transfer
		26.1.3 Fresh vs. Frozen
	26.2 Embryo Assessment, Selection and Decision-Making
		26.2.1 Do Not Disturb (DND)
		26.2.2 Timelapse and CareMaps
		26.2.3 PGT-A
		26.2.4 The Future (Artificial Intelligence)
	26.3 The Endometrium
	26.4 Training and Competency of Embryo Transfer Personnel
	26.5 Problematic Embryo Transfers
		26.5.1 Reloading
	26.6 Embryo Transfer Catheters
	26.7 Laboratory Process
		26.7.1 Standard Incubation
		26.7.2 CareMaps
		26.7.3 Embryo Transfer Preparation
		26.7.4 Hyaluronan Enriched Media
	References
27 Data and Patient Management in the IVF Laboratory
	27.1 Introduction
		27.1.1 The Past
		27.1.2 The Present
		27.1.3 The (Near) Future
	27.2 Digitisation in the IVF Clinic
		27.2.1 Information Databases
		27.2.2 Consents
		27.2.3 The Laboratory
		27.2.4 Cyber Security
		27.2.5 Quality Management
	27.3 Conclusion
28 Genomics and Genetic Counselling: Considerations and Challenges with IVF Patients
	28.1 Introduction
		28.1.1 Genetics and Genomics: An Overview
		28.1.2 Genetic Counselling
	28.2 Genetic Causes of Infertility
	28.3 Options Available to Individuals Wishing to Reduce Risk of Passing a Genetic Condition to their Child
	28.4 Preimplantation Genetic Testing
	28.5 General Considerations for Clinical Scientists Supporting Patients in PGT Treatment Pathways
	28.6 Gamete Donation
	28.7 Other Considerations for Genetics/Genomics for Clinical Scientists Working in the ART Laboratory
		28.7.1 Extended Carrier Screening (ECS)
		28.7.2 Direct-to-Consumer Testing for Carrier Screening
	28.8 Futures
		28.8.1 Polygenic Risks Scores (PRS)
		28.8.2 Non-Invasive Preimplantation Genetic Testing of Embryos
		28.8.3 Metabolomics and Proteomics
		28.8.4 Genome Editing
	References
29 Third Party Reproduction
	29.1 Introduction
	29.2 Management of Donation in IVF Clinics in the UK
	29.3 HFEA (Human Fertilisation and Embryology Authority) Regulatory Requirements and Clinic Policy for Donation and Surrogacy
	29.4 Gamete Donation
	29.5 Embryo Donation
	29.6 Donor Expenses
	29.7 Anonymity for Donors
	29.8 Donor Screening, Including Intended Parents in a Surrogacy Arrangement
	29.9 Family Limit
	29.10 Matching Donors and Recipients
	29.11 Carrier Screening
	29.12 Egg Donation
	29.13 Egg Sharing
	29.14 Sperm Donation
	29.15 Shared Motherhood/Parenthood
	29.16 Egg and Sperm Recipients
	29.17 Egg Recipients
		29.17.1 Fresh Donation Treatment
		29.17.2 Frozen Egg Donation Treatment
	29.18 Surrogacy
	29.19 Legal Parenthood in Surrogacy
	29.20 National Health Service (NHS) Funding for Donation Services
	29.21 Fertility Tourism
	References
30 Add-Ons: Critical Evaluation of Supplementary Treatments
	30.1 Time Lapse Imaging Systems (TLS)
	30.2 Hyaluronate Enriched Medium
	30.3 Assisted Hatching (AH)
	30.4 Artificial Oocyte Activation (AOA)
	30.5 Pre-Implantation Genetic Testing for Aneuploidy (PGT-A)
	30.6 Endometrial Receptivity Analysis (ERA)
	30.7 Future Assessment of Add-ons
	References
31 Cryobiology and Cryopreservation of Oocytes and Embryos
	31.1 History of Gamete and Embryo Cryopreservation
	31.2 Principles of Cryobiology
	31.3 Cryoprotectants Used in Cryomedia
	31.4 Oocyte Cryopreservation
	31.5 Embryo Cryopreservation
	31.6 Developments in Cryopreservation Materials and Methods
		31.6.1 Automated Vitrification
		31.6.2 Artificial Intelligence (AI)
		31.6.3 In Vitro Maturation
		31.6.4 Ovarian Tissue Cryopreservation
		31.6.5 Re-vitrification/Warm Cycles
	31.7 Management of Gametes and Embryos Cryotanks
	31.8 Clinical Frozen Embryo Transfers (FET) Outcomes
	References
32 In Vitro Maturation (IVM) and Artificial Oocyte Activation (AOA)
	32.1 In Vitro Maturation
	32.2 Artificial Oocyte Activation
	32.3 Conclusion
	References
33 ICSI: Micromanipulation Setup, Consumables, and Operational Procedure
	33.1 History of ICSI
	33.2 Indications for ICSI
	33.3 ICSI Procedure
		33.3.1 Oocyte Denudation
		33.3.2 Dish Preparation
		33.3.3 Sperm Selection and Immobilisation
		33.3.4 Sperm Injection
	33.4 Developments in ICSI
		33.4.1 Artificial Oocyte Activation (AOA)
		33.4.2 Modified ICSI (mICSI)
		33.4.3 Physiological Intracytoplasmic Sperm Injection (PICSI)
	33.5 Factors Affecting Success Rates of ICSI Procedure
		33.5.1 Polar Body Positioning
		33.5.2 Timing of Denudation and Injection, Including IVM and Rescue ICSI
		33.5.3 Troubleshooting
	33.6 Risks, Success Rates and Long-Term Health of Offspring
	References
34 Embryo Biopsy and Preimplantation Genetic Testing (PGT)
	34.1 Introduction
	34.2 The Embryo Culture Environment
	34.3 Embryo Biopsy
		34.3.1 Polar Body Biopsy
		34.3.2 Cleavage Stage Biopsy
		34.3.3 Morula Biopsy
		34.3.4 Trophectoderm Biopsy
		34.3.5 Sample Preparation and Tubing
		34.3.6 Pre-Implantation Genetic Testing
		34.3.7 Adverse Outcomes and Misdiagnosis
	References
35 Computer Assisted Sperm Analysis (CASA)
	35.1 Introduction
	35.2 Principles of CASA Operation
		35.2.1 Count and Concentration
		35.2.2 Motility
		35.2.3 Quality Control
		35.2.4 Morphology
		35.2.5 CASA and IVF
	35.3 Conclusion
	References
36 Time-Lapse Embryo Culture Systems and Morphokinetics in Clinical Embryology: A Brief Overview on the Evolution of Embryo Scoring System
	36.1 Description of a Time-Lapse System (TLS)
	36.2 Oocyte Morphology
	36.3 Fertilisation Assessment
	36.4 Early Embryo Development
	36.5 Compaction to Blastulation
		36.5.1 What to Annotate
	36.6 Quality Assured Manual Annotations
	36.7 Automated Annotations
	36.8 Use and Development of Prediction Algorithms
	36.9 Benefits of Morphokinetics and TL Culture for the Patient
	36.10 Conclusion
	References
37 Abnormal Cleavage, Self-Correction of Aneuploidy and Decision-Making Regarding Embryo Fate
	37.1 The Cell Cycle
	37.2 Abnormal Patterns of Cell Division
		37.2.1 Direct Cleavage/Multichotomous Mitosis
		37.2.2 Rapid Cleavage
		37.2.3 Reverse Cleavage (RC)/Cell Fusion
		37.2.4 Fragment Generation/Reabsorption
		37.2.5 Exclusion of Cells from the Morula
	37.3 Preferential Allocation of Aneuploid Cells to the Trophectoderm Model
	37.4 Self-Correction: Where Are We Now?
	37.5 Decision-Making Regarding Embryo Fate
	37.6 Conclusion
	References
38 Artificial Intelligence Application in the ART Laboratory
	References
Index