Freezing Test in Gujarat | Female First Hospital

It is possible to cryopreserve sperm and embryo at -196º C in liquid N2 by computerized slow cooling method. Subsequent cryothaw cycles treatment becomes cost effective and patient does not require undergoing active cycle management. Embryos and sperms can be kept cryopreserved for years.Recently developed vitrification technique uses rapid cooling of embryo.

A. Embryo

B. Semen

C. Oocyte

A. Cryopreservation and Vitrification of Embryo

Egg retrieval under ultrasound guidance and subsequent fertilization and embryo culture are carried out according to our current procedures. If there happens to be a surplus of embryos following selection for fresh transfer (usually between one to four embryos are transferred to the uterus), then embryos of sufficient quality may be considered for cryostorage. While embryos can be frozen at any preimplantation stage between one-cell (one day old) to the blastocyst stage (5-6 days old). In certain cases where all embryos need to be frozen without a fresh transfer (e.g., when a woman may be at risk from ovarian hyperstimulation that might be complicated by pregnancy), we generally freeze all embryos the day after egg collection at the one-cell stage.

Techniques of controlled-rate freezing are utilized that slowly cool embryos in cryoprotectant fluid (“anti-freeze” solution) from body temperature down to -196°C, at which temperature they are stored in containers of liquid nitrogen called dewars. The embryos are actually contained within special indelibly labeled plastic vials, or straws, that are sealed prior to freezing. Once frozen, they are placed inside labeled tubes attached to aluminum cans and stored in numbered canisters within the liquid nitrogen dewar. Site and label designations are stored in three separate file systems to avoid confusion and misidentification of cryopreserved embryos. When it comes time to thaw the embryos, all available identifiers of the stored specimen must match and be confirmed before thawing commences. The embryos are thawed out at room temperature, which takes about one to two minutes. However, the most critical element of the thaw procedure is not the timing but the careful dilution of the cryoprotectant fluid to return the embryo to its favored culture medium. This permits resumed growth and development in vitro. Once this is done, the embryo is assessed for cryodamage to determine if it is suitable for transfer. Experience has shown that if the embryo survives 50% or more intact, it is worthwhile to replace it. Embryos can accommodate such levels of cellular damage and still establish healthy pregnancies.

Varying strategies may be applied according to how many and which embryos are thawed prior to transfer. It should be noted that not every couple undergoing IVF will need to worry about embryo freezing/thawing, since not every couple will have sufficiently large number of “surplus” or non-transferred embryos available for freezing. In the event that there are more than two or three embryos frozen, thawing is usually undertaken until two to three healthy appearing embryos are recovered. In some cases, this may mean that all the cryopreserved embryos are thawed, in others just two or three. There always remains a possibility that there may be no embryo survival after thaw occurs, and no transfer is possible. If many early embryos are frozen, it is possible to thaw all of them and culture them for several days to allow selection of the best for transfer. When too many embryos are available for transfer in this circumstance, then extra embryos of sufficient quality may be refrozen for later use. This course of action has produced healthy offspring, proving the efficacy of double freezing of embryos.

During a medication-prepared frozen/thawed embryo transfer cycle as a patient, you will follow a treatment schedule using Lupron, estrogen (pills) and progesterone (suppository) in order to achieve appropriate endometrium (uterine wall lining) for embryo transfer. Following embryo transfer, estrogen and progesterone will be administered daily until the 7th to 8th week of pregnancy or until a negative pregnancy test.

Considerations and Risk

The Ethics Committee of the American Society of Reproductive Medicine (ASRM) has published guidelines for ethical consideration of human embryo cryopreservation. Possible advantages of cryopreservation of embryos suggested by the Committee include

• Reduction of the risk of triplets or quadruplets by cryopreservation of embryos exceeding an optimal number for transfer to an individual patient.
• Possibly increasing pregnancy rates by replacing thawed embryos during spontaneous ovulatory cycles or cycles in which the estrogen and progesterone hormone levels do not exceed that which occurs naturally.
• Possibly decreasing the number of stimulated ovary drug treatment cycles needed for the attainment of pregnancy.

The primary concern with the use of cryopreservation techniques is the possible loss of embryos to cryoinjury.,.

Another concern with cryopreservation is the potential risk of birth defects in children produced from frozen/thawed embryos. In the domestic animal industry, large-scale freezing and transfer of embryos has not resulted in increased birth defects. Studies to date on those human offspring arising from thawed embryos have not shown any significant increase in abnormalities when compared to pregnancy outcomes in the rest of the population.

When a specimen is processed for cryopreservation:

• A semen analysis is performed on each ejaculate. This includes a complete seminal fluid analysis quantitating sperm motility, forward progression, sperm density, and morphology.
• All specimens are stored in liquid nitrogen storage tanks (-196_C).
• There does not appear to be any increased risk of birth defects using frozen semen.
• There is no guarantee that any given specimen will necessarily produce a pregnancy; however, recent studies indicate that the overall success rate following the use of cryopreserved semen is from 40%-50%.

embryos are available for transfer in this circumstance, then extra embryos of sufficient quality may be refrozen for later use. This course of action has produced healthy offspring, proving the efficacy of double freezing of embryos.

During a medication-prepared frozen/thawed embryo transfer cycle as a patient, you will follow a treatment schedule using Lupron, estrogen (pills) and progesterone (suppository) in order to achieve appropriate endometrium (uterine wall lining) for embryo transfer. Following embryo transfer, estrogen and progesterone will be administered daily until the 7th to 8th week of pregnancy or until a negative pregnancy test.

Semen freezing & Oocyte Freezing

Egg Freezing Program Candidacy

Recent advances in the In Vitro Fertilization process have now allowed us to successfully freeze, store and later thaw and fertilize cryopreserved human eggs. This new advance in the field of assisted reproductive technologies provides yet another valuable tool to aid in the management of couples struggling to overcome problems with infertility. While we have enjoyed excellent pregnancy rates for some time utilizing frozen and later thawed human embryos, our new ability to store unfertilized human eggs allows many new techniques and resources to be developed.

In addition, our increasing ability to successfully cryopreserve oocytes allows us to now offer the option of oocyte freezing and storage to women concerned about preserving their own fertility. This may be a viable option for women facing premature loss of their ovaries or their ovarian function or for women concerned about advancing age and it’s effect on their reproductive potential. Harvesting and cryopreservation of eggs may also be considered by women dealing with cervical, endometrial and breast cancer, as well as those facing systemic chemotherapy or other therapy thought to be a threat to reproductive potential.

Group “A” Candidates

Group “B+” Candidates

• Good chances for success
• Ages 35-37 years.
• Serum FSH “fertility thermostat” value less than 8.1 mIU/ml on cycle day 3. Satisfactory ovarian reserve testing. If being treated for cancer, short term, limited exposure to radiation or chemotherapy with no direct radiation of ovaries.

Group “C” Candidates

• Moderate chances for success.
• Ages 37.5-39 years
• Serum FSH “fertility thermostat” value between 7.1 and 9.0 mIU/ml on cycle day 3. Satisfactory ovarian reserve testing. If being treated for cancer, short term, limited exposure to radiation or chemotherapy with no direct radiation of ovaries.

Group “C-” Candidates

• Some guarded chance of success.
• Ages 39.1 to 40 years.
• Serum FSH “fertility thermostat” between 5.0 and 8.5 mIU/ml on cycle day 3. Satisfactory ovarian reserve testing. If being treated for cancer, no more than short term, limited exposure to radiation or chemotherapy with no direct radiation of ovaries.
• Non-Eligible Candidates
• Age over 40 years or any woman with unsatisfactory ovarian reserve testing outcomes.
• Serum FSH “fertility thermostat” levels greater than 10.0 mIU/ml on cycle day 3 If being treated for cancer, extended chemotherapy or direct radiation of the ovaries. You can find other programs that will freeze your eggs at thisage, but there are NO reports from anywhere in the world that such freezing offers a reasonable chance for success.

Steps in the Egg Freezing Process

Our treatment protocol involves the preparation of your ovaries for the production of the mature eggs needed for the freezing process. Full details of this process will be provided at the time of your formal entry into the program, but a summary of the process is provided here for your reference and understanding:

Step-1: Normal Menstruation

Involves waiting for your normal menstrual period to begin. Depending upon the treatment protocol prescribed for you, with the beginning of your menses, you will either be given instructions to begin your fertility drugs, or instructions to wait for a certain date within the three weeks following the start of your period to begin a medication to allow the Doctors the opportunity to optimize your ovaries for the fertility drugs that will follow.

Step-2: Ovarian Stimulation

Involves the administration of fertility medications designed to allow your ovaries to begin the growth of the several eggs that will be removed for subsequent freezing. During this phase, you will be self-administering daily fertility injections to allow for the successful production of multiple healthy eggs. You will be shown how to take these injectable medications prior to using them. While taking these medications, you will need to be seen for painless ultrasound studies and blood tests 3-4 times over the 10-12 day period these medicines are being used. These studies may be carried out near your home, or at any of our offices.

Step-3: Egg Retrieval

After your eggs have been determined to have matured adequately, you will be scheduled for the surgical harvesting of your eggs. This is done by the Doctor at our facilities. In nearly all instances, the procedure is carried out with a light sedative and pain medication that allows for a very rapid recovery. You will be discharged from the office when it has been determined that you are stable, usually within 1-3 hours of your procedure.

Step-4: Oocyte Cryopreservation

After your eggs have been recovered, they are prepared in our cryopreservation laboratory by the embryologist and who will assure the healthy appearance of the eggs and begin the process of preparing the eggs for freezing. We use the most advanced “vitrification” (snap freezing) techniques for egg freezing that are providing success rates far superior to earlier “slow freezing” methods. The freezing process rapidly lowers the temperature of human eggs to a point where all metabolic processes (aging) is halted.

Step 5: Oocyte Storage

Following the freezing process, your eggs will be transferred to a liquid nitrogen storage chamber where they are able to be effectively stored in the frozen state. The initial, short term storage of your eggs will occur at our facility, however long term storage will be required at a special “cryobank” which is a facility where long term secure maintenance of frozen human specimens occurs. You will be given information concerning a contract that you will need to enter into providing for the long-term storage of your eggs.

Step-6: Thaw and Use of Your Frozen Eggs

When the time arrives for you to use your cryopreserved eggs, you will need to make arrangements with our facility at least 3 months in advance of the time you desire the eggs to be thawed and fertilized. It must be remembered that the eggs have been frozen UNFERTILIZED, and that the fertilization of previously frozen eggs requires special techniques and handling. The successful fertilization of cryopreserved eggs is greatly enhanced by the use of a process called “ICSI” (intrascreeningcytoplasmic sperm injection). Utilizing this method assures the best chance for the fertilization of the eggs and their subsequent development into healthy embryos with the potential to produce an ongoing pregnancy. When we are contacted by you with a request for use of the frozen eggs, we will begin making the arrangements for the treatment cycle required to assure the best chance for a successful pregnancy outcome for you.

The Use of Egg Freezing to Pause the Biological Clock

For younger women seeking to guard and extend their fertility, egg freezing offers perhaps the greatest advance in fertility preservation in the history of modern medicine. Every day, modern informed, forward looking women are helping assure their future fertility potential at our center. Unlike many unproven programs offering egg freezing as a new venture, the Fertility Institutes have many, many healthy babies born following oocyte cryopreservation.