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EMBRYO
CULTURE
What is en embryo culture
• Embryo culture is a component of in
vitro fertilisation where in resultant embryos
are allowed...
• Embryo culture can be performed in two ways
Artificial medium
autologous endometrial coculture
• Artificial culture medi...
 Single or sequential medium are equally
effective for the culture of human embryos to
the blastocyst stage.
 Artificial...
SINGLE CULTURE
• the same culture medium throughout the
period
SEQUENTIAL
CULTURE
• the embryo is sequentially placed in d...
COCULTURE
• Autologous Endometrial Coculture is a
technique of assisted reproductive technology.
It involves placing a pat...
How Coculture is performed
Patient undergoes an endometrial biopsy during
which a small piece of her uterine lining is
rem...
• Once fertilization is confirmed, the
patient’s embryos are placed on top of her
own (and now thawed) endometrial cells.
...
The potential candidate
• Coculture can be an effective treatment for
patients who have failed previous IVF cycles or
who ...
EGG RETRIEVING
• Egg is
TECHNIQUES IN EMBRYO
CULTURE
Oocyte Wash Buffer
• On the day of egg retrieval (Day 0), this buffer
is used for the retrieval of the eggs from the
ovary...
Fertilization Medium
• After the wash at retrieval, the eggs are put into
the fertilization medium. This medium contains a...
Cleavage Medium
• All of the eggs which undergo normal
fertilization are next placed into cleavage
medium, which is formul...
Blastocyst Medium
• Embryos, that are to be cultured until Day 5 or 6,
are placed, later on Day 3, into another medium
ref...
Sperm Buffer
• The sperm buffer is formulated in order to
maintain the correct pH when the solution is
exposed to air. Thi...
Sperm Medium
• The sperm medium is similar to the Sperm
Buffer except that the buffer is such that the
correct pH of the s...
BLASTOCYST CULTURE
BLASTOCYS
T
An embryo that has developed for
five to seven days after fertilization
and has 2 distinct cell types and a
ce...
Blastocyst development
• The blastocyst usually forms on day 5 as fluid builds
within the compacted morula
• A healthy bla...
Blastocyst formation rate
• The goal of in vitro fertilization and embryo culture is
to provide high quality embryos which...
Advantages of blastocyst transfer for
IVF
• One problem with this is that 2 to 3-day-old embryos are
normally n the fallop...
How Long Should an
Embryo Be Cultured?
• Two Days: Embryos that are cultured for two
days are generally transferred at the two or
four-cell stage. This type of t...
Three Days
• : Embryos that are cultured
for three days are usually
transferred at the six to eight
cell stage. Many labor...
Five Days: Embryos that are cultured for five days
are transferred at the blastocyst stage. Blastocysts
consist of 12 to 1...
The Embryo Culture Equipment
The Laminar Flow Hood
• The preparation of all media and solutions to
be used in IVF, ICSI and IUI occurs inside this
spec...
The Preparation Incubator
• All dishes and solutions to be used for an IVF,
ICSI or IUI treatment are maintained in this
i...
Embryo Culture Incubator
• All eggs and embryos are incubated here
throughout their time in the VFC laboratory. The
unit i...
IVF Chamber
• Whenever the eggs and embryos need to be
outside of the incubator for any reason, they
are handled in our IV...
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Embryo culture

Embryo culture is a component of in vitro fertilisation where in resultant embryos are allowed to grow for some time in an artificial medium before being inserted into the uterus.

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Embryo culture

  1. 1. EMBRYO CULTURE
  2. 2. What is en embryo culture • Embryo culture is a component of in vitro fertilisation where in resultant embryos are allowed to grow for some time in an artificial medium before being inserted into the uterus. •
  3. 3. • Embryo culture can be performed in two ways Artificial medium autologous endometrial coculture • Artificial culture medium can be single culture sequential culture
  4. 4.  Single or sequential medium are equally effective for the culture of human embryos to the blastocyst stage.  Artificial embryo culture media contains- glucose, pyruvate, and energy-providing components  Amino acids, nucleotides, vitamins, and cholesterol improve the performance of embryonic growth and development
  5. 5. SINGLE CULTURE • the same culture medium throughout the period SEQUENTIAL CULTURE • the embryo is sequentially placed in different media
  6. 6. COCULTURE • Autologous Endometrial Coculture is a technique of assisted reproductive technology. It involves placing a patient’s fertilized eggs on top of a layer of cells from her own uterine lining, creating a more natural environment for embryo development and maximizing the chance for an in vitro fertilization (IVF) pregnancy.
  7. 7. How Coculture is performed Patient undergoes an endometrial biopsy during which a small piece of her uterine lining is removed. The uterine lining sample is sent to a research lab, where it is treated, purified and frozen. The patient then undergoes a typical IVF cycle and is given medication to stimulate egg growth in her ovaries. The patient’s eggs are retrieved and mixed with the sperm. At this time, the lab begins thawing and growing her endometrial cells.
  8. 8. • Once fertilization is confirmed, the patient’s embryos are placed on top of her own (and now thawed) endometrial cells. Over the next two days, the embryos are closely monitored for growth and development. The patient’s embryos are transferred into her uterus for implantation and pregnancy
  9. 9. The potential candidate • Coculture can be an effective treatment for patients who have failed previous IVF cycles or who have poor embryo quality.
  10. 10. EGG RETRIEVING • Egg is
  11. 11. TECHNIQUES IN EMBRYO CULTURE
  12. 12. Oocyte Wash Buffer • On the day of egg retrieval (Day 0), this buffer is used for the retrieval of the eggs from the ovary. Oocyte wash buffer has an ingredient, which prevents a change in pH when the solution is exposed to air during the retrieval. The eggs are very susceptible to any minute changes in the pH of their environment. The eggs are washed in this buffer and then placed into the next medium for culture.
  13. 13. Fertilization Medium • After the wash at retrieval, the eggs are put into the fertilization medium. This medium contains a variety of salts, sugars, amino acids, protein and other nutrients essential for the maintenance of the egg (and sperm in IVF) during the process of fertilization (IVF and ICSI). The fertilization medium and all of the other subsequent culture media, are buffered with the appropriate components in order to maintain the correct pH of the solution in the embryo incubator.
  14. 14. Cleavage Medium • All of the eggs which undergo normal fertilization are next placed into cleavage medium, which is formulated specifically to support the growth requirements of the early cleavage stage embryo. The cleaving (dividing) embryo is cultured in this medium until Day 3. If the embryo transfer is scheduled for Day 3, the embryos are transferred to the uterus in a small amount of this medium.
  15. 15. Blastocyst Medium • Embryos, that are to be cultured until Day 5 or 6, are placed, later on Day 3, into another medium referred to as blastocyst medium. The embryos are then maintained in this medium until embryo transfer on Day 5 or embryo cryopreservation on Day 5 or 6. This medium has additional components and/or different components required by the embryo in its transition from a cleavage stage embryo to a blastocyst. If the embryo transfer is scheduled on Day 5, the embryos are transferred to the uterus in a small amount of this medium.
  16. 16. Sperm Buffer • The sperm buffer is formulated in order to maintain the correct pH when the solution is exposed to air. This buffer is used during the preparation of semen samples and solutions for semen samples, which will be washed and processed outside of the incubator.
  17. 17. Sperm Medium • The sperm medium is similar to the Sperm Buffer except that the buffer is such that the correct pH of the solution is maintained whilst in the incubator. This medium is important for the final resuspension of sperm to be used in IVF because the process of fertilization occurs inside the incubator.
  18. 18. BLASTOCYST CULTURE
  19. 19. BLASTOCYS T An embryo that has developed for five to seven days after fertilization and has 2 distinct cell types and a central cavity filled with fluid (blastocoel cavity) The cells in a blastocyst have just started to differentiate The surface cells that surround the cavity (just under the outer shell) are called the trophectoderm and will later develop into the placenta A more centrally located group of cells - the inner cell mass, will become the fetus
  20. 20. Blastocyst development • The blastocyst usually forms on day 5 as fluid builds within the compacted morula • A healthy blastocyst often begins hatching from its outer shell, called the zona pellucida between day 5 to day 7 after fertilization • Within 24 hours after hatching, embryo implantation after IVF (or a "natural" pregnancy) begins as the embryo invades into the uterine lining • The blastocyst releases HCG hormone (the pregnancy test hormone) which leaks into the mother's blood as the embryo implants
  21. 21. Blastocyst formation rate • The goal of in vitro fertilization and embryo culture is to provide high quality embryos which are capable of continued development and result in live births • However, under standard IVF culture conditions, only about 25 to 60% of human embryos progress to the blastocyst stage after 5 days of culture • The low rate of embryo development has 2 main causes: – A less than optimal culture environment in the lab dish – The inherent "weakness" of human embryos • Therefore, in the past embryos were transferred to the uterus after 2 or 3 days
  22. 22. Advantages of blastocyst transfer for IVF • One problem with this is that 2 to 3-day-old embryos are normally n the fallopian tubes, not in the uterus. The embryo gets to the uterus about 80 hours after ovulation. • Embryo implantation process begins about 3 days later - after blastocyst formation and hatching out of the embryonic shell have occurred. • Therefore, if in vitro culture conditions are maximized so healthy blastocysts form at a high rate, then day 5 blastocyst embryo transfer can be done. – The uterine lining on day 5 should be receptive to the arriving embryo - this a more "natural" time for the embryos to be in the uterus. It is the same timing as with a natural pregnancy. – The transfer is done shortly before the time for actual invasion and implantation
  23. 23. How Long Should an Embryo Be Cultured?
  24. 24. • Two Days: Embryos that are cultured for two days are generally transferred at the two or four-cell stage. This type of transfer is beneficial for couples who have a low number of embryos available for transfer, or who have embryos that are developing poorly.
  25. 25. Three Days • : Embryos that are cultured for three days are usually transferred at the six to eight cell stage. Many laboratories prefer to culture embryos until this stage because it allows for increased monitoring. Embryos cultured for three days can be checked by the embryologist for gene activation and cleavage, which improves the likelihood of transferring a viable embryo.
  26. 26. Five Days: Embryos that are cultured for five days are transferred at the blastocyst stage. Blastocysts consist of 12 to 16 cells and are well on their way to be ready for implantation into the uterus Many labs opt to transfer at the blastocyst stage, particularly if you have had repeated miscarriages or IVF failures.
  27. 27. The Embryo Culture Equipment
  28. 28. The Laminar Flow Hood • The preparation of all media and solutions to be used in IVF, ICSI and IUI occurs inside this specialized hood, which blows air out towards the embryologist. The air is filtered and the outflow of air prevents any contaminants from blowing in and contaminating the solutions and embryo dishes being prepared. Preparation of semen samples to be used in IVF, ICSI and IUI also occurs in this sterile environment.
  29. 29. The Preparation Incubator • All dishes and solutions to be used for an IVF, ICSI or IUI treatment are maintained in this incubator until use. The incubator is sterile inside, is at 37°C, has a carbon dioxide concentration of 6.0%, and the environment is fully humidified to prevent any evaporation. All solutions and dishes to be used for treatment are equilibrated in this incubator for a minimum of 4 hours before use.
  30. 30. Embryo Culture Incubator • All eggs and embryos are incubated here throughout their time in the VFC laboratory. The unit is infused with the proper levels of oxygen and carbon dioxide to ensure that the eggs/embryos are maintained under optimum conditions at all times. The environment in the incubator is also humidified and kept at 37°C. The temperature and gas levels are monitored continuously and the incubator is attached to a telephone based alarm system which will call out to the embryologist during off hours should an unsuitable or emergency condition arise.
  31. 31. IVF Chamber • Whenever the eggs and embryos need to be outside of the incubator for any reason, they are handled in our IVF Chamber. The chamber looks like an isolate that you would see in a special care newborn nursery in the hospital. This chamber however is specially modified and adapted for the purpose of maintaining eggs and embryos under optimum conditions even when they are being handled outside of the incubator.

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