embryology

1. FACTORS AFFECTING EMBRYO
QUALITY
Dr. Pratixa Joshi
Senior Clinical Embryologist
Director and Chairperson
“MAA” Multispecialty Hospital and IVF Center

4. The Culture System
• Contact materials/toxicity
• Group embryo culture vs. individual
• Incubator type/management
• Low O2 vs. atmospheric O2
• Air quality/VOCs
• Technician

5. Factors affecting Oocytes Quality In Vitro
Oocytes are Sensitive to temperature alterations
Meiotic spindle disruption, Increased aneuploidy
other adverse affects on oocytes (Pickerning et al .,1990,Wang et al 2001)
 All disposables and media at 37C
 Calibrated heating devices
Temperature control during follicular aspiration,
During transport of aspirate to the work station,
During agg search and handling of COCs
During ICSI

6.  Homeostasis
Protect from loss of amino acids :use properly formulated flush/wash media
 Oocytes (ph:7.4) are sensitive to extracellular pH fluctuations
Cytoskeletal and metabolic changes
Constant pH of 7.3 in media
HEPES buffered media (CO2 independent)
Bicarbonate buffered media (CO2 dependant)
 Oocytes are also sensitive to changes in Osmolarity
Oocyte shrinking/sweling
Cover media with oil
Factors affecting Oocytes Quality In Vitro

8. Osmolality
• Proper media osmolality ~260-290mOsm
• >300mOsm can inhibit embryo development in vitro
(Hadi et al. 2005)
• Lab technique can inadvertently raise media osmolality
(Swain et al. 2012)

9. Thermal control during ICSI
FERTILITY AND STERILITY Vol 77.No:6June :2006

10. Temperature

13. Effects of Pathological Levels of ROS during IVF

16. Zygote Morphology and Embryo Cleavage scores
related to the women Age

17. Pregnancy and Delivery rates according to the Age

18. Fertility and Sterility® Vol. 98, No. 1, July 2012
0015-0282/$36.00

19. Association Between Body Mass Index and Oocyte and
Embryo Characteristics

22. • Patient Selection
• a. Age
b. Diagnosis; male and female
c. Ovarian reserve
d. Smoking
e. Other pre-IVF evaluations; HSG, SIS, anti-sperm antibodies,
acrosome reaction
• Stimulation Protocols
• a. General Stimulation Flow Sheet
b. Long vs. Short Lupron
c. Antagonist
d. FSH vs. HMG
e. Premature Luteinization
f. Fluid in cavity during stimulation

24. • Factors affecting IVF outcome
• There is no doubt that IVF helps to diagnose problems of fertilization and embryo development that could not be assessed by any other
method. It is both a diagnostic and therapeutic procedure. The complexity of human reproduction means that many factors may play a
role in the success or failure of IVF.
• In order to gain some understanding of the factors that may affect the outcome of IVF, I will try and to dissect the IVF procedure to see
which factors are present which may affect outcome.
• Patient Selection
• a. Age
b. Diagnosis; male and female
c. Ovarian reserve
d. Smoking
e. Other pre-IVF evaluations; HSG, SIS, anti-sperm antibodies, acrosome reaction
• Stimulation Protocols
• a. General Stimulation Flow Sheet
b. Long vs. Short Lupron
c. Antagonist
d. FSH vs. HMG
e. Premature Luteinization
f. Fluid in cavity during stimulation
• Egg and Embryo Factors
• a. Number of eggs
b. Fragmentation
c. Embryo growth
d. Immature eggs at retrieval
e. Fractured zona
f. Polyspermy
• Embryo Transfer Technique
• a. Day 3 transfer
b. Blastocyst transfer
c. Trial transfer
• Immune Factors
• Sperm Factors
• a. Balanced translocations.
b. Y-Chromosome micro deletions.
c. Cystic Fibrosis mutations.
d. DNA fragmentation
• Factors in the Lab
• a. Air quality
b. Temperature