CRYOPRESERVATION
EGG FREEZING

Oocyte Cryopreservation(OC) is a process by which eggs are formed, extracted and stored for
future use in
a lady.
The lady undergoes a transvaginal ultrasound, evaluation of hormones and detailed counseling
about the
prospects in her particular case prior to starting the cycle.
EGG FREEZING PROCESS TIMELINE
Day 2 -patient reports for basal scan and blood tests.

Gonadotropin injections are started on a daily basis.

Day 5-transvaginal ultrasound monitoring started.

Antagonist injection started on days 5-7 depending on scan findings.

Day 9-12-Trigger injection given once follicles reach mature size.

Ovum pick up done under anaesthesia

Collected mature eggs are frozen in liquid nitrogen for future use.

Result time!
FAQ
OC was originally introduced as a fertility preservation option for those women
who were going to undergo surgical removal of the ovaries or chemotherapy
treatment for cancer.
Here it is usually done as an emergency procedure as treatment for cancer will
need to be initiated as soon as possible.
It is the same process of OC done as a planned procedure where the woman wants to
preserve her eggs for social rather than medical reasons.
The better terminology proposed is OC for anticipated gamete exhaustion(AGE)
Unfortunately, the reproductive window is definitely shorter in women than in
men.
The time period when fertility rates peak in a lady is also the same as when her
education and career advancement are at a peak.
It gives women more
reproductive choice and decreases the pressure they face from the “biological
clock”
NO, the best option will always be less invasive methods and natural conception
if it is a possibility.
It is important to counsel women that though OC is an option to consider, it can
in no manner be looked at as an option which will provide a 100% guarantee of
giving birth.
The chances of a successful outcome primarily increase with two factors;
A.The younger the age at which the eggs are frozen- women who freeze their eggs
at less than 35 years of age have more chances of giving birth.
B.The more eggs that are frozen, the better the chances of pregnancy.
One latest study reported that if a woman less than 35 years freezes 10-20 eggs,
she has a 70-90% chance of having at least one child from them.
At age 38 and above, a woman may need to freeze about 20-30 eggs to have a
reasonable chance of having at least one live child.
This may increase the number of cycles needed to collect the same number of eggs
as the age of the lady increases.
The main complication is risk of OHSS as our aim is deliberate hyperstimulation
to get the maximum number of eggs possible.
The risk of OHSS can be minimized by ;
A. Using the antagonist protocol.
B. Using the agonist trigger and avoiding hCG trigger.
C. Using antagonist injections post ovum pick up.
With the advent of the vitrification procedure, the rates of survival after
thawing have increased substantially.
Rates of fertilization, implantation and clinical pregnancy rates have been
reported to be similar for both fresh and frozen oocytes.
But, the success rates may also differ with the expertise and experience of the
person who does the vitrification.
As yet, there have been no increased risk of congenital abnormalities reported in
children born from frozen eggs as compared to other IVF pregnancies.
But the data on long-term oocyte storage and long-term offspring health will be
known only with time and increased use of the treatment.
FREEZING OF SPERM
The semen sample of the male partner is processed to remove the debris and separate the fastest moving sperms which are then frozen in liquid nitrogen for future use.
The semen sample of the male partner is processed to remove the debris and separate the fastest moving sperms which are then frozen in liquid nitrogen for future use.

- In case of sperm donation in semen banks.
- In men who have difficulty collecting samples at the time of IUI due to more stress and pressure to provide “on-demand”.
- In men who are going to undergo treatment for cancer which may destroy their sperms.
- As a routine in IVF cycles to have a backup sample in case the husband is not available on the date of pick up or is unable to collect on that day.
- As a safety precaution in men whose counts are subnormal, when they decide to wait or try other alternative treatment options in the hope that the counts may improve
- In couples where the husband is stationed overseas for long periods but they want to continue treatment with a few cycles of IUI.
- In all cases where sperm has been surgically retrieved as in after PESA, TESA or TESE.
SPERM FREEZING PROCESS TIMELINE
Husband or the male partner is tested for sexually transmissible diseases like HbsAg, HIV and VDRL

Asked to come to the lab with a sex-free or ejaculation free interval of 3 to 5 days.

Informed consent is taken.

Semen sample is collected by masturbation.

The sample is processed by the embryologist.

The processed sample is mixed with cryoprotectant and frozen in liquid nitrogen.
FAQ
It is essential to do a serology test before going ahead with freezing the semen
sample.
He should be free from any active urogenital infection
Semen should be collected by masturbating into a wide-mouthed sterile container. Due to concerns with chances of infection, it should not be collected by sex and withdrawal method.
This is a fairly common problem as it is not always possible to masturbate on
demand, especially in a bathroom or hospital setting where there is a lack of
privacy.
Aids such as porn magazines, porn videos, use of a vibrator, or partner-assisted
masturbation in a private room may be tried.
If it still fails to give results, we advise the couple to bring the sample from
home provided the hospital is reachable within 30 min of collection.
In such cases, an additional consent stating that the sample brought from home
is the husband's sample should be signed by both the partners.
There is no set time limit for the storage of sperm but the usual time frame
stipulated is a period of ten years.
Sperms have been reported to have been used successfully even 40 years after
storage!
Semen parameters prior to freezing will determine the parameters post
thawing.
Sperm may undergo some damage during the freezing and thawing process due to
osmotic or oxidative stress as well as the formation of intracellular ice
crystals, which in turn may reduce the number of functionally normal sperms.
The process of cryopreservation must hence be carefully controlled to minimize
the risk of damage.
One recent study reported that about 80% of sperms may survive the thaw.
Sperm can be stored for the purpose of donation only in ART banks registered
under the provisions of this act.
Sperm can be donated only by a male in the age group of 21 to 55 years.
It is stored after testing the person for diseases outlined in the
guidelines.
Sperm donated by one person can be used for only one commissioning couple in a
lifetime and not more.
The number of samples we need to freeze will vary from person to person depending
upon the reason for which it is done, the semen parameters, the treatment
proposed as well as family building goals of the couple.
One semen sample may give up to four vials of sperm of counts normal.
One vial is used for one IUI attempt typically.
But for IVF/ICSI as even one vial may contain millions of sperms,even one vial
should be sufficient.
The usual time period we collect a sample is a sex-free or ejaculation-free gap of 3 to 5 days, so samples can be collected with a gap of few days between them. But having said that, it can be collected at shorter intervals also if time is of concern, as in starting treatment for cancer.
EMBRYO CRYOPRESERVATION

This is the process of freezing embryos by a process called vitrification for intended long-term use in the future.
Why is it done?
1. When excess embryos are made in an IVF cycle, in order to avoid the risk of OHSS, we may choose to freeze
them all and replace them in subsequent months.
2. As typically only 2-3 embryos maximum are transferred at one shot, the excess embryos, if any, will be
frozen for future use with the consent of the couple.
3. For couples requiring embryo donation.
4. For couples who are married and want to freeze their embryos as they want to postpone childbearing for
personal reasons.
5. When embryos need to be tested for any genetic diseases prior to using them.
What is the process?
The formation of embryos is by an IVF/ICSI cycle give link to the same

Once embryos are formed, the excess is frozen by a process called vitrification.
FAQ
Embryo freezing has been in common use since the 1980s and has been proven to be
a safe and effective method, especially with the advent of vitrification.
There is a theoretical risk of embryos getting damaged or not surviving when
thawed.,
As usually only good quality embryos are chosen to be frozen,the risk of
damaging the embryos remains negligible in experienced hands.
There is no theoretical limit to the number of years where they remain viable but under the latest Indian law, storage is allowed for a period of ten years.
YES, World over there has been an increase in the number of frozen embryo
transfers(FET) in the past decade.
Frozen embryo transfers are reported to give as good as and in many instances
better pregnancy rates when compared to fresh transfers.
In case you have finished your family or do not want to renew storage for some
other reason, you can choose to,
1. Discard the embryos.
2. Donate them for research or for training purposes.
This is usually covered in the consent form signed at the start of the process
itself.
YES, Informed consent is taken from both partners before proceeding with
freezing.
Consent is also taken about what to do in case there is the untimely death of
one partner or in case of legal divorce.