Delhi Public School, Noida

Biology CBSE Project

Submitted By: Chhavi Singh

Xii- Science
 Acknowledgements…

I would like to pay my heartfelt thanks to my subject teacher

__________________________________ for guiding me throughout this tenure.

I would also like to thank my family and friends for helping me in

this project.
 Certificate
This is to certify that the Project work has been satisfactorily
completed under my guidance and supervision by Ms. Chhavi
Singh, student of XIIth Science.

T. Signature

______________
 Index

 Topic
 Definition & Meaning
 Brief Discussion
 Examples
 Detailed study of In-vitro fertilization
 Method
 Success Rates
 Expansion of IVF
 Risks
 Bibliography
 Topic :
Assisted
Reproductive Technologies
(Case Study)
 Definition & Meaning

Assisted reproductive technology (ART) a general term referring to methods

used to achieve pregnancy by artificial or partially artificial means. It is reproductive
technology used primarily in infertility treatments. Some forms of ART are also used in fertile
couples for genetic reasons. ART is also used in couples who are discordant for certain
communicable diseases, e.g. AIDS, to reduce the risk of infection when a pregnancy is
desired. Examples of ART include in vitro fertilisation, intra-cytoplasmic sperm injection
(ICSI), cryopreservation, and intrauterine insemination (IUI). There is yet no strict definition
of the term. Usage of the ART mainly belongs in the field of reproductive endocrinology and
infertility.
Definition:

The Centres for Disease Control and Prevention (CDC)—which is required as a

result of the 1992 Fertility Clinic Success Rate and Certification Act to publish the annual
ART success rates at U.S. fertility clinics—defines ART to include "all fertility treatments in
which both eggs and sperm are handled. In general, ART procedures involve surgically
removing eggs from a woman's ovaries, combining them with sperm in the laboratory, and
returning them to the woman's body or donating them to another woman." According to
CDC, "they do not include treatments in which only sperm are handled (i.e., intrauterine—or
artificial—insemination) or procedures in which a woman takes medicine only to stimulate
egg production without the intention of having eggs retrieved."
 Brief Discussion
ART has been used in the United States since 1981 to help women become pregnant, most
commonly through the transfer of fertilized human eggs into a woman’s uterus (in vitro
fertilization). However, deciding whether to undergo this expensive and time-consuming
treatment can be difficult.

According to CDC’s 2010 ART Success Rates, 147,260* ART cycles were performed at 443
reporting clinics in the United States during 2010, resulting in 47,090 live births (deliveries of
one or more living infants) and 61,564 infants. Although the use of ART is still relatively rare
as compared to the potential demand, its use has doubled over the past decade. Today, over
1% of all infants born in the U.S. every year are conceived using ART.

*Excludes cycles in which a new treatment procedure was being evaluated.

The National Institute of Child Health and Human Development held a
workshop on September 12-13, 2005, to summarize the risks for adverse pregnancy outcomes
after assisted reproductive technology (ART), develop an approach to counseling couples
regarding these risks, and establish a research agenda. Although the majority of ART children
are normal, there are concerns about the increased risk for adverse pregnancy outcomes.
More than 30% of ART pregnancies are twins or higher-order multiple gestations (triplets or
greater) and more than one half of all ART neonates are the products of multifetal gestations,
with an attendant increase in prematurity complications. Assisted reproductive technology
singleton pregnancies also demonstrate increased rates of perinatal complications-small for
gestational age infants, preterm delivery, and perinatal mortality-as well as maternal
complications, such as preeclampsia, gestational diabetes, placenta previa, placental
abruption, and cesarean delivery. Although it is not possible to separate ART-related risks
from those secondary to the underlying reproductive pathology, the overall increased
frequency of obstetric complications, including preterm birth and small for gestational age
neonates, should be discussed with the couple. Significant gaps in knowledge were identified,
and the basic science and clinical and epidemiologic research required to address these gaps
is outlined.
 Examples
 Artificial Insemination
 Cloning
 Embryonic splitting or cleavage
 Cryopreservation of sperms, oocytes or embryos
 Embryo transfer
 Fertility medication
 Hormone treatment
 In-vitro fertilization
 Intra-cytoplasmic sperm injection
 Pre-implantation genetic diagnosis
 Artificial Insemination

Artificial insemination (AI) is the deliberate introduction of semen into a female for the
purpose of fertilisation, by means other than ejaculation directly into the vagina or oviduct.

Artificial insemination is a fertility treatment for humans, and is a common practice in the
breeding of dairy cattle and pigs. Artificial insemination may employ assisted reproductive
technology, donated sperm, and/or animal husbandry techniques.

In Humans
Artificial insemination is a means for a woman to conceive when she does not have a male
partner, when she does not want a male partner, or when a male partner's sexual inhibition or
physical limitation impedes his ability to impregnate her by sexual intercourse. Women who
have issues with the cervix such as cervical scarring, cervical blockage from endometriosis,
or thick cervical mucus also benefit from AI since the sperm must pass through the cervix to
result in fertilization.
 Cloning

Human cloning is the creation of a genetically identical copy of a human. It does not refer
to monozygotic multiple births or the reproduction of humans/animals cells or tissue. The
ethics of cloning is an extremely controversial issue. The term is generally used to refer to
artificial human cloning; human clones in the form of identical twins are commonplace, with
their cloning occurring during the natural process of reproduction.

There are two commonly discussed types of human cloning: therapeutic

cloning and reproductive cloning. Therapeutic cloning involves cloning cells from an adult
for use in medicine and transplants, and is an active area of research. Reproductive cloning
would involve making cloned humans, for couples wanting to have a child, but cannot
naturally.
 Embryonic splitting or
cleavage

In embryology, cleavage is the division of cells in the early embryo. The zygotes of many
species undergo rapid cell cycles with no significant growth, producing a cluster of cells the
same size as the original zygote. The different cells derived from cleavage are called
blastomeres and form a compact mass called the morula. Cleavage ends with the formation of
the blastula.

Depending mostly on the amount of yolk in the egg, the cleavage can be holoblastic (total or
entire cleavage) or meroblastic (partial cleavage). The pole of the egg with the highest
concentration of yolk is referred to as the vegetal pole while the opposite is referred to as
the animal pole.

Cleavage differs from other forms of cell division in that it increases the number of cells
without increasing the mass. This means that with each successive subdivision, the ratio of
nuclear to cytoplasmic material increases.
 Cryopreservation of sperms, oocytes or
embryos

Cryopreservation is a process where sperms, oocytes and embryos are preserved by cooling
to sub-zero temperatures, typically 77 K (= −196 °C, the boiling point of liquid nitrogen). At
these cold temperatures, any biological activity, including the biochemical reactions that
would cause cell death, is effectively stopped. However, if cryoprotectant solutions are not
used, the cells being preserved are likely to be damaged due to freezing during the cooling or
thawing process.
 Embryo Transfer

Embryo transfer refers to a step in the process of assisted reproduction in which embryos
are placed into the uterus of a female with the intent to establish a pregnancy. This technique
(which is often used in connection with in vitro fertilization (IVF)), may be used in humans
or in animals, in which situations the goals may vary.
 Fertility Medication

Fertility medication are drugs which enhance reproductive fertility. For women, fertility
medication is used to stimulate follicle development of the ovary. There are currently very
few fertility medication options available for men.
 Hormone Treatment

Hormone treatment or hormonal therapy is the use of hormones in medical

treatment. Treatment with hormone antagonists may also referred to as hormonal therapy.
 In-vitro fertilization

In vitro fertilization (IVF) is the technique of letting fertilization of the male and female
gametes (sperm and egg) occur outside the female body.

Techniques usually used in in vitro fertilization include:

 Transvaginal ovum retrieval (OCR) is the process whereby a small needle is inserted
through the back of the vagina and guided via ultrasound into the ovarian follicles to
collect the fluid that contains the eggs.
 Embryo transfer is the step in the process whereby one or several embryos are placed into
the uterus of the female with the intent to establish a pregnancy
 Intra-cytoplasmic sperm
injection

Intracytoplasmic sperm injection (ICSI) is a procedure in which a single sperm is injected

directly into an egg.

This procedure is most commonly used to overcome male infertility problems, although it
may also be used where eggs cannot easily be penetrated by sperm, and occasionally in
addition tosperm donation.

It can be used in teratozoospermia, because once the egg is fertilized, abnormal sperm
morphology does not appear to influence blastocyst development or blastocyst
[3]
morphology. Even with severe teratozoospermia, microscopy can still detect the few sperm
cells that have a "normal" morphology, allowing for optimal success rate.
 pre-implantation genetic
diagnosis

In medicine and (clinical) genetics pre-implantation genetic diagnosis (PGD or PIGD) (also
known as embryo screening) refers to procedures that are performed on embryos prior
toimplantation, sometimes even on oocytes prior to fertilization. PGD is considered another
way to prenatal diagnosis. When used to screen for a specific genetic disease, its main
advantage is that it avoids selective pregnancy termination as the method makes it highly
likely that the baby will be free of the disease under consideration. PGD thus is an adjunct
to assisted reproductive technology, and requires in vitro fertilization (IVF) to
obtain oocytes or embryos for evaluation.
 Detailed study of In-vitro
fertilization
In vitro fertilisation (IVF) is a process by which an egg is fertilised by sperm outside the
body: in vitro. IVF is a major treatment for infertility when other methods of assisted
reproductive technology have failed. The process involves monitoring a woman's ovulatory
process, removing ovum or ova(egg or eggs) from the woman's ovaries and
letting sperm fertilize them in a fluid medium in a laboratory. When a woman's natural cycle
is monitored to collect a naturally selected ovum (egg) for fertilisation, it is known as natural
cycle IVF. The fertilised egg (zygote) is then transferred to the patient'suterus with the
intention of establishing a successful pregnancy. The first successful birth of a "test tube
baby", Louise Brown, occurred in 1978. Louise Brown was born as a result of natural cycle
IVF. Robert G. Edwards, the physiologist who developed the treatment, was awarded
the Nobel Prize in Physiology or Medicine in 2010.
The term in vitro, from the Latin meaning in glass, is used, because early biological
experiments involving cultivation of tissues outside the living organism from which they
came, were carried out in glass containers such as beakers, test tubes, or petri dishes. Today,
the term in vitro is used to refer to any biological procedure that is performed outside the
organism it would normally be occurring in, to distinguish it from an in vivo procedure,
where the tissue remains inside the living organism within which it is normally found. A
colloquial term for babies conceived as the result of IVF, "test tube babies", refers to the
tube-shaped containers of glass or plastic resin, called test tubes, that are commonly used in
chemistry labs and biology labs. However, in vitro fertilisation is usually performed in the
shallower containers called Petri dishes. One IVF method, Autologous Endometrial
Coculture, is actually performed on organic material, but is still considered in vitro.

IVF may be used to overcome female infertility in the woman due to problems of
the fallopian tube, making fertilisation in vivo difficult. It may also assist in male infertility,
where there is a defect in sperm quality, and in such cases intracytoplasmic sperm
injection (ICSI) may be used, where a sperm cell is injected directly into the egg cell. This is
used when sperm have difficulty penetrating the egg, and in these cases the partner's or a
donor's sperm may be used. ICSI is also used when sperm numbers are very low. ICSI results
in success rates equal to those of IVF.

For IVF to be successful it typically requires healthy ova, sperm that can fertilise, and
a uterus that can maintain a pregnancy. Due to the costs of the procedure, IVF is generally
attempted only after less expensive options have failed.

IVF can also be used with egg donation or surrogacy where the woman providing the egg
isn't the same who will carry the pregnancy to term. This means that IVF can be used for
females who have already gone through menopause. The donated oocyte can be fertilised in
a crucible. If the fertilisation is successful, the embryo will be transferred into the uterus,
within which it may implant.

IVF can also be combined with preimplantation genetic diagnosis (PGD) to rule out presence
of genetic disorders. A similar but more general test has been developed
called Preimplantation Genetic Haplotyping
 Method
Theoretically, in vitro fertilisation could be performed by collecting the contents from a
woman's fallopian tubes or uterus after natural ovulation, mixing it with semen, and
reinserting into the uterus. However, without additional techniques, the chances of pregnancy
would be extremely small. Such additional techniques that are routinely used in IVF
include ovarian hyper stimulation to retrieve multiple eggs, ultrasound-guided transvaginal
oocyte retrieval directly from the ovaries, egg and sperm preparation, as well as culture and
selection of resultant embryos before embryo transfer back into the uterus.
 Egg & Sperm Preparation

In the laboratory, the identified eggs are stripped of surrounding cells and prepared
for fertilisation. An oocyte selection may be performed prior to fertilisation to select eggs
with optimal chances of successful pregnancy. In the meantime, semen is prepared for
fertilisation by removing inactive cells and seminal fluid in a process called sperm washing.
If semen is being provided by a sperm donor, it will usually have been prepared for treatment
before being frozen and quarantined, and it will be thawed ready for use.
 Fertilization
The sperm and the egg are incubated together at a ratio of about 75,000:1 in the culture
media for about 18 hours. In most cases, the egg will be fertilised by that time and the
fertilised egg will show two pronuclei. In certain situations, such as low sperm count or
motility, a single sperm may be injected directly into the egg using intracytoplasmic sperm
injection (ICSI). The fertilised egg is passed to a special growth medium and left for about 48
hours until the egg consists of six to eight cells.

In gamete intrafallopian transfer, eggs are removed from the woman and placed in one of the
fallopian tubes, along with the man's sperm. This allows fertilisation to take place inside the
woman's body. Therefore, this variation is actually an in vivo fertilisation, not an in
vitro fertilisation.
 Embryo Culture

Typically, embryos are cultured until having reached the 6–8 cell stage three days after
retrieval. In many Canadian, American and Australian programmes, however, embryos are

placed into an extended culture system with a transfer done at the blastocyst stage at around
five days after retrieval, especially if many good-quality embryos are still available on day 3.
Blastocyst stage transfers have been shown to result in higher pregnancy rates.[9] In Europe,
transfers after 2 days are common.

Culture of embryos can either be performed in an artificial culture medium or in

an autologous endometrial coculture (on top of a layer of cells from the woman's own uterine
lining). With artificial culture medium, there can either be the same culture medium
throughout the period, or a sequential system can be used, in which the embryo is
sequentially placed in different media. For example, when culturing to the blastocyst stage,
one medium may be used for culture to day 3, and a second medium is used for culture
thereafter. Single or sequential medium are equally effective for the culture of human
embryos to the blastocyst stage. Artificial embryo culture media basically contain glucose,
pyruvate, and energy-providing components, but the addition of amino acids, nucleotides,
vitamins, and cholesterol improve the performance of embryonic growth and
development. Methods to permit dynamic embryo culture with fluid flow and embryo
movement are also available. A new method in development uses the uterus as an incubator
and the naturally occurring intrauterine fluids as culture medium by encapsulating the
embryos in permeable intrauterine vessel.
 Embryo Selection & transfer
Laboratories have developed grading methods to judge oocyte and embryo quality. In order
to optimise pregnancy rates, there is significant evidence that a morphological scoring system
is the best strategy for the selection of embryos. However, presence of soluble HLA-G might
be considered as a second parameter if a choice has to be made between embryos of
morphologically equal quality. Also, two-pronuclear zygotes (2PN) transitioning through
1PN or 3PN states tend to develop into poorer-quality embryos than those that constantly
remain 2PN.

Embryos are failed by the embryologist based on the amount of cells, evenness of growth
and degree of fragmentation. The number to be transferred depends on the number available,
the age of the woman and other health and diagnostic factors. In countries such as Canada,
the UK, Australia and New Zealand, a maximum of two embryos are transferred except in
unusual circumstances. In the UK and according to HFEA regulations, a woman over 40 may
have up to three embryos transferred, whereas in the USA, younger women may have many
embryos transferred based on individual fertility diagnosis. Most clinics and country
regulatory bodies seek to minimise the risk of pregnancies carrying multiples, as it is not
uncommon for more implantations to take than desired. The embryos judged to be the "best"
are transferred to the patient's uterus through a thin, plastic catheter, which goes through
her vagina and cervix. Several embryos may be passed into the uterus to improve chances
of implantation and pregnancy.
 Success Rates
IVF success rates are the percentage of all IVF procedures which result in a favorable
outcome. Depending on the type of calculation used, this outcome may represent the number
of confirmed pregnancies, called the pregnancy rate or number of live births, called the live
birth rate.

Due to advancement in reproductive technology, the IVF success rates are substantially better
today than they were just a few years ago. The most current data available in the United
States a 2009 summary complied by the Society for Reproductive Medicine which reports the
average national IVF success rates per age group using non-donor eggs.
 Expansions of IVF
In zygote intrafallopian transfer (ZIFT), egg cells are removed from the woman's ovaries and
fertilized in the laboratory; the resulting zygote is then placed into the fallopian tube. The
following are techniques generally requires methods of in vitro fertilisation. In vitro
fertilization, however, usually does not require these techniques.

 Assisted zona hatching (AZH) is performed shortly before the embryo is transferred to the uterus.
A small opening is made in the outer layer surrounding the egg in order to help the embryo hatch
out and aid in the implantation process of the growing embryo.

 Intracytoplasmic sperm injection (ICSI) is beneficial in the case of male factor infertility
where sperm counts are very low or failed fertilization occurred with previous IVF
attempt(s). The ICSI procedure involves a single sperm carefully injected into the center
of an egg using a microneedle. This method is also sometimes employed when donor
sperm is used.

 Autologous endometrial coculture is a possible treatment for patients who have failed previous
IVF attempts or who have poor embryo quality. The patient's fertilized eggs are placed on top of a
layer of cells from the patient's own uterine lining, creating a more natural environment for
embryo development.

 Cytoplasmic transfer is the technique in which the contents of a fertile egg from a donor are
injected into the infertile egg of the patient along with the sperm.
 Egg donors are resources for women with no eggs due to surgery, chemotherapy, or genetic
causes; or with poor egg quality, previously unsuccessful IVF cycles or advanced maternal age.
In the egg donor process, eggs are retrieved from a donor's ovaries, fertilized in the laboratory
with the sperm from the recipient's partner, and the resulting healthy embryos are returned to the
recipient's uterus.
 Sperm donation may provide the source for the sperm used in IVF procedures where the male
partner produces no sperm or has an inheritable disease, or where the woman being treated has no
male partner.
 A gestational carrier is an option when a patient's medical condition prevents a safe
pregnancy, when a patient has ovaries but no uterus due to congenital absence or
previous surgical removal, and where a patient has no ovaries and is also unable to carry
a pregnancy to full term.
 Preimplantation genetic diagnosis (PGD) involves the use of genetic screening
mechanisms such as Fluorescent In Situ Hybridization (FISH) or Comparative Genomic
Hybridization (CGH) to help identify genetically abnormal embryos and improve healthy
outcomes.

 Embryo splitting can be used for twinning to increase the number of available embryos.
 Risks
The risk of birth defects in infants born following assisted reproductive technology (ART)
treatment is a controversial question. Most publications examining the prevalence of birth
defects in ICSI and IVF infants compared to spontaneously conceived infants have serious
methodological limitations; despite this, most researchers have concluded that there is no
increased risk. METHODS: We carried out a systematic review to identify all papers
published by March 2003 with data relating to the prevalence of birth defects in infants
conceived following IVF and/or ICSI compared with spontaneously conceived infants.
Independent expert reviewers used criteria defined a priori to determine whether studies were
suitable for inclusion in a meta-analysis. Fixed effects meta-analysis was performed for all
studies and reviewer-selected studies. RESULTS: Twenty-five studies were identified for
review. Two-thirds of these showed a 25% or greater increased risk of birth defects in ART
infants. The results of meta-analyses of the seven reviewer-selected studies and of all 25
studies suggest a statistically significant 30–40% increased risk of birth defects associated
with ART.

CONCLUSIONS: Pooled results from all suitable published studies suggest that children
born following ART are at increased risk of birth defects compared with spontaneous
conceptions. This information should be made available to couples seeking ART treatment.
 Bibliography

www.google.com
www.wikipedia.com
Various magazines and newspapers