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Biology
Third Year of Secondary School

Explainer: Artificial Reproduction

In this explainer, we will learn how to describe the processes of test-tube baby formation (IVF) and
renucleation and outline the role of gamete banks in artificial reproduction.

In 2006, Dr. Patricia Rashbrook (born in 1943) gave birth to a healthy baby boy. Dr. Rashbrook and
the father of her child were in their 60s when they had this child, and biologically, this seems to not
make sense. Menopause, the point in a woman’s life when egg production stops, is typically reached
around the age of 51. So, how did this couple manage to conceive a healthy new baby in their 60s?

This baby was born through in vitro fertilization (IVF), which is a form of artificial reproduction that
allows egg and sperm cells to be joined in a laboratory environment, in order to help with infertility.
The word in vitro means “in glass” and is meant to describe how experiments can be done outside
the living organism, as in the glass of a test tube or a plastic petri dish.

Artificial, or assisted, reproduction is the creation of new life by artificial means. By combining the
egg and sperm in the lab, the resulting embryo can be transferred into the uterus where implantation
and pregnancy occur.

The term test-tube baby is sometimes used when referring to IVF, but it can be misleading and
inappropriate for some people. The term was first coined because of the idea that an embryo is formed
using laboratory equipment instead of being formed in the woman’s fallopian tube. This is only true
for the very early stages of an embryo’s development because the embryo is eventually transferred
into the uterus for embryo implantation and pregnancy. But as the term was being used casually in
public, it created a false impression that the whole pregnancy takes place outside the uterus. For that
reason, many countries lean toward using the term in vitro fertilization to refer to the process instead.

Key Term: Artificial Reproduction

Artificial reproduction is the creation of new life in ways that are not natural.

Key Term: In Vitro Fertilization (IVF)

In vitro fertilization (IVF) is when an egg is fertilized by a sperm in a petri dish in a laboratory, usually
to allow parents to reproduce, where they would otherwise be unable to.

IVF is a process with multiple stages.

The first stage involves stimulation with hormones, that can be synthetic, in order to promote egg
development in the ovaries. Typically, only one egg develops and undergoes ovulation, and in the

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early stages of IVF, this one egg is what is extracted. However, the use of hormones has allowed
multiple eggs to develop simultaneously in a process called “controlled ovarian hyperstimulation.”

Key Term: Ovulation

Ovulation is the part of the menstrual cycle when an egg is released from one of a woman’s ovaries.

Next, the eggs, or oocytes, can be extracted in a clinical setting in what is sometimes called an “egg
retrieval” or “oocyte pick up.” While the eggs are being extracted, the sperm can be prepared from
the ejaculated semen.

Then, the eggs and sperm are combined for fertilization. Normally, this stage occurs in the fallopian
tubes, but in IVF, this occurs in the IVF lab. Typically, a petri dish is used, which is a plastic (or glass)
dish where the eggs and sperm can be deposited. They are suspended in a specialized liquid culture
media that is meant to mimic the physiological environment of the fallopian tube. You can see this
in the photograph below.

Figure 1

By combining the egg and sperm together, the sperm will bind and penetrate the egg to fertilize it
over time.
Once the eggs and sperm are combined, they are placed in incubators that maintain the same pH and
temperature found in a fallopian tube environment where they normally grow. After fertilization, the
next stage involves monitoring the resulting embryos over the course of about 3–5 days before they
can be transferred to the uterus. There are different stages of an embryo’s development, day 3 is called
the cleavage stage, day 4 is called the morula stage, and day 5 is called the blastocyst stage.
During the final stage of IVF, the embryo is loaded into a specialized catheter that is inserted into
the uterus. The embryo is then transferred into the uterus where it will hopefully implant and lead
to a pregnancy. Any excess embryos created during an IVF procedure can be frozen and transferred
at another time.
In some cases, women may transfer more than one embryo. This can lead to an increased chance of
having fraternal, or nonidentical, twins.

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The basic steps of IVF are demonstrated in Figure 2.

(1) Hormones are taken to stimulate

the ovaries

(3) Sperm is extracted

(2) Eggs are extracted from the mother
from the father

(4) The eggs and sperm are combined in

fertilization outside of the body

(5) Embryos are allowed to develop for a

short period of time

(6) Embryos are then transferred back into

the uterus

Figure 2: A diagram that shows the basic outline of in vitro fertilization (IVF).

Example 1: Describing the Stages of IVF

Where are embryos placed in the final stage of the IVF process?

A. Uterus
B. Cervix
C. Ovary
D. Vagina
E. Fallopian tube (oviduct)

Answer

Infertility can impact many couples around the world. In vitro fertilization (IVF) is a technique that
allows the eggs and sperm to be combined outside of the body in order to overcome some of the
causes of infertility.

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IVF has multiple stages.

First, the female is treated with hormones, which can be synthetic, to develop multiple eggs in the
ovaries. These eggs can then be extracted and brought to the IVF lab along with the sperm.

Then, the eggs and sperm are combined in a petri dish and are allowed to fertilize. A petri dish is a
plastic or glass dish that contains a specialized culture media fluid to mimic the environment of the
fallopian tubes and encourage fertilization. Once the eggs and sperm are combined, they are placed
in special incubators to maintain specific pH and temperature levels that are optimal for fertilization
and embryo growth.

After fertilization, the resulting embryos grow for around 3–5 days and are transferred back to the
uterus. Here, they will hopefully implant in the uterine lining and lead to a pregnancy.

Therefore, the correct answer is A: uterus.

IVF success rates are mostly dependent on maternal age, which may have to do with decreases in egg
quality over time. According to the Centers for Disease Control and Prevention in the USA in 2016,
women under the age of 35 have the highest IVF pregnancy rates at about 30–40%. Meaning that out
of 100 women (aged less than 35) who undergo IVF, about 30 to 40 of them will get pregnant. The
pregnancy rate then begins to decline to less than 5% by the age of 40. Note that these are statistics
for infertile women undergoing IVF, while the rest of the population (without diagnosed infertility)
may have higher pregnancy rates even with advanced age.
Due to the decline in pregnancy rates with increased age, couples who are planning to delay
parenthood are encouraged to freeze their gametes at an earlier age. This is important because IVF
can be very expensive, and for some older couples, they may not be able to afford the multiple rounds
of IVF needed to get pregnant, so they save their gametes when they are young.
Gametes are also encouraged to be frozen and cryogenically stored for those preparing for
chemotherapy, as chemotherapy may reduce the viability or quality of the gametes. These gametes
can be stored under liquid nitrogen at very cold temperatures in egg banks or sperm banks
(collectively gamete banks) for up to 10 years and more. You can see an example of samples stored
in a part of a cryogenic storage tank in the photograph below.

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 Figure 3

Besides humans, gametes of animals can also be preserved by cryopreservation. This can be helpful
for endangered animals that may soon become extinct.

Key Term: Cryopreservation

Cryopreservation is a process that enables the long-term conservation of biological material by

cooling at very low temperature (typically −196∘ C when using liquid nitrogen). At this very low
temperature, enzymatic and chemical reactions that could damage the biological material are
stopped.

Key Term: Gamete Banks

Gamete banks are storage facilities that specialize in storing cryopreserved gametes like eggs or
sperm.

Example 2: Describing Gamete Banks

What is stored at very cold temperatures in gamete banks?

A. Fertilized eggs
B. Embryos
C. Sperm only

D. Unfertilized eggs and sperm

E. Unfertilized eggs only

Answer

Artificial reproduction is the creation of new life by artificial means. One prominent example is
in vitro fertilization (IVF), where eggs and sperm are removed from the body and combined in a
laboratory to form embryos that can then be transferred into the uterus to create a pregnancy. Excess
embryos can be frozen in liquid nitrogen for future use.

Gametes, such as unfertilized eggs and sperm, can also be frozen in egg banks or sperm banks
(collectively called gamete banks), and this has some advantages. For example, somebody who needs
to undergo chemotherapy may want to freeze their gametes since the treatment may damage their
gametes.

Let’s look at the different answers and see which one best defines what is stored in gamete banks.

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In A, “fertilized eggs” are not stored in gamete banks because fertilized eggs are not gametes. A
fertilized egg is a zygote.

In B, “embryos” are not stored in gamete banks because, again, they are not gametes.

In C, “sperm only” is half true because sperm can be stored in gamete banks but so can eggs.

In D, “unfertilized eggs and sperm” seem like the best choice because both of these are gametes and
can both be stored in gamete banks.

In E, “unfertilized eggs only” again is only half true because sperm can also be stored in gamete
banks.

Therefore, the correct answer is D: unfertilized eggs and sperm.

Besides IVF, another popular technique is artificial insemination, where sperm is transferred to the
reproductive tract in the hopes of starting a pregnancy. In one type of artificial insemination, called
intrauterine insemination (IUI), sperm is transferred directly to the uterus. These techniques are
used in humans and are a very popular method to inseminate farm animals.

Definition: Artificial Insemination

Artificial insemination is the process of transferring sperm directly into the female reproductive tract
to achieve pregnancy.
A popular form of beef comes from the Japanese black cattle breed “Wagyu.” These cattle have a
high concentration of fat inside their muscle tissue that gives the meat a lower melting point and a
very tender quality unmatched by other cattle. Because of the high demand for Wagyu, breeders can
use IVF or artificial insemination to make more of these cattle at a lower cost.
Breeders are able to make many Wagyu embryos from a single cow by first stimulating the cow with
hormones to produce multiple eggs in the ovaries. Then, the eggs can either be extracted and mixed
with sperm for IVF or the sperm can be transferred directly into the uterus by artificial insemination.
The resulting Wagyu embryos can be harvested and then transferred into non-Wagyu cows, which are
much cheaper than Wagyu cows. This way breeders are able to make hundreds of Wagyu offspring
quickly and for less money.
Other advantages of using artificial insemination or IVF in cattle include the ability to improve the
pregnancy rates in breeds with low fertility, to have better control over crossbreeding to create new
desirable breeds (for example, through crossing cattle with disease resistance and milk production),
or to choose the gender of the offspring.
You will recall that female somatic cells typically have two X chromosomes and males have one X
and one Y chromosome. The egg cell always contains a single X chromosome, while the sperm may
contain either an X or Y chromosome. Therefore, when the haploid sperm and egg combine to form
the diploid embryo, it is the sperm that determines the gender. The same is true in cattle (and most
mammals).
The X chromosome contains more DNA than the Y chromosome, and because of this difference, it
can be separated using centrifugation and electric fields as well as a technique called flow cytometry.

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Once separated, the sperm can be frozen and used by farmers to manage the gender of their cattle.
So, if they need more female cows for milk production for one particular season, they could use
the sperm with the X chromosome to inseminate the females by artificial insemination. This will
produce female embryos. This process produces what is known as sexed semen, because it can be
used to produce offspring of a desired gender.

Example 3: Using Artificial Reproduction for Gender Selection

Sperm samples from farm animals can be separated into X- and Y-chromosome-containing sperm
cells and stored in liquid nitrogen until needed. In which situation might this procedure be beneficial
to farmers?

A. To prioritize meat production in sheep by using sperm cells containing X chromosomes

B. To prioritize milk production in sheep by using sperm cells containing Y chromosomes

C. To prioritize meat production in cattle by using sperm cells containing X chromosomes

D. To prioritize milk production in cattle by using sperm cells containing Y chromosomes
E. To prioritize milk production in cattle by using sperm cells containing X chromosomes

Answer

Artificial reproduction is the creation of new life by artificial means. Artificial reproduction in farm
animals can be done using in vitro fertilization (IVF) or artificial insemination.

In vitro fertilization (IVF) is a form of artificial reproduction where eggs and sperm are removed
from the body and combined in a laboratory to form embryos that can then be transferred into the
uterus to lead to a pregnancy. Artificial insemination is the transfer of sperm directly into the female
reproductive tract that will lead to a pregnancy.

In mammals, female somatic cells have two copies of the X chromosome, while male cells have
one X chromosome and one Y chromosome. Therefore, the female gamete (the egg cell) can only
contain a single X chromosome, and the male sperm contains either the X chromosome or the Y
chromosome. So, when the haploid sperm and egg combine to form the diploid embryo, it is the
sperm that determines the biological sex.

The X chromosome is larger than the Y chromosome and can be separated based on its size. Once
separated, the appropriate sperm can be used in IVF or artificial insemination to breed animals of a
specific biological sex.

Let’s look at these different answers to see which is beneficial for farmers.

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In A, the answer is “to prioritize meat production in sheep by using sperm cells containing X
chromosomes.” Sheep, when farmed for meat, can be either female or male, so this answer is not
a benefit.

In B, the answer is “to prioritize milk production in sheep by using sperm cells containing Y
chromosomes.” The Y chromosome in sperm would make male sheep, which do not produce milk
(only the females do), so this would not be a benefit.

In C, the answer is “to prioritize meat production in cattle by using sperm cells containing X
chromosomes.” Cattle, when farmed for meat, can be either female or male. Male cattle are more
frequently used however, so there may be a benefit in choosing sperm with a Y chromosome and not
an X chromosome.

In D, the answer is “to prioritize milk production in cattle by using sperm cells containing Y
chromosomes.” Female cattle produce milk, and not males, so using a Y chromosome here would
not help because only males would be produced.

In E, the answer is “to prioritize milk production in cattle by using sperm cells containing X
chromosomes.” This would produce female cattle that would make milk, so this would be a benefit
for the farmer.

Therefore, the correct answer is E.

The manipulation of gametes in a laboratory setting has led to other technologies besides IVF.
Suppose you wanted to clone, or make a genetic copy of, a particular breed of cattle; could you use
the cells in a steak to do this?

Key Term: Clone

A clone is a genetically identical copy of an organism or DNA sequence. DNA cloning is the process
of making more copies of a particular segment of DNA.
Yes, you can! All the genetic information to create a particular breed of cattle is contained in the
nuclei of the body cells—or somatic cells—of an individual cow or bull. As meat products, like steak,
are mostly the muscle tissue of an animal, we can take the nuclei from these muscle cells! To make
a clone of this DNA, a technique called somatic cell nuclear transfer (or renucleation) can be done.
This involves several steps.
First, a cell is taken from the steak and its nucleus is isolated.
Then, the nucleus from a cow’s egg cell is removed or enucleated.
The steak cell nucleus and enucleated cow egg are then combined by transferring the nucleus into
the enucleated egg.
This can then be transferred to a cow where it can implant to form an offspring that is genetically
identical to the cow that the steak was produced from. This is the process they used to clone a
particular breed of cattle called Delta in 2014 and can be seen in Figure 4.

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 Key Term: Enucleation

Enucleation is the process of removing a nucleus from a cell.

Key Term: Somatic Cell Nuclear Transfer (Renucleation)

Somatic cell nuclear transfer is a specialized technique used to clone an animal. It involves replacing
the egg nucleus with the nucleus of a body (somatic) cell of a particular animal to create a clone of
that animal.

Extract body
(somatic) cell
Isolate nucleus Somatic cell
nuclear transfer

Extract egg Remove egg

cell nucleus

Figure 4: An illustration showing somatic cell nuclear transfer.

This process was also done with frogs. Embryonic cells at varying stages of development were
transferred into enucleated frog eggs. These frogs then developed as clones of the original embryos
from which they were derived.
This process of cloning using somatic cell nuclear transfer differs from sexual reproduction to
produce an offspring. In sexual reproduction, a haploid egg cell (𝑛) is fertilized with a haploid sperm
cell (𝑛) to produce a genetically unique diploid embryo (2𝑛) that is a combination of the genetics
from the egg and sperm. In somatic cell nuclear transfer, the nucleus of a somatic diploid body cell
(2𝑛) of the individual or embryo is transferred into an enucleated egg cell to produce an embryo
that is clone of the organism from which the nucleus is derived. This process can be used in other
organisms besides cattle, including frogs and sheep for example.

Example 4: Understanding the Technique of Somatic Cell Nuclear Transfer to Create

Clones

The two sheep in the figure shown were used in a somatic cell nuclear transfer procedure. Eggs were
obtained from sheep 1, and skin cells were obtained from sheep 2.

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 Sheep 1 Sheep 2

What would the genetic makeup of the offspring produced by this process be?

A. The offspring would be genetically identical to sheep 2.

B. The offspring would be genetically identical to sheep 1.

C. The offspring would be genetically different to sheep 1 and sheep 2.

D. The offspring would be completely black in color.
E. The offspring would be completely white in color.

Answer

Artificial reproduction is the creation of new life by artificial means. One prominent example is
in vitro fertilization (IVF), where eggs and sperm are removed from the body and combined in a
laboratory to form embryos that can then be transferred into the uterus to create a pregnancy.

Another form of artificial reproduction is called somatic cell nuclear transfer or renucleation. This
is a technique that can be used to clone, or make genetic copies, of animals.

There are multiple steps to clone, so let’s explain this in the context of this question.

First, the skin cell nucleus of sheep 2 is isolated.

Then, the nucleus from the egg cell of sheep 1 is removed or enucleated.

Then, the skin cell nucleus of sheep 2 and the enucleated sheep 1 egg are combined by transferring
the nucleus into the enucleated egg.

The result is an embryo that can then be transferred to a sheep where it can implant to form an
offspring that is genetically identical to sheep 2. You can see an overview of this process below.

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 Extract body
(somatic) cell
Isolate nucleus
Somatic cell
nuclear transfer
Sheep 2

Extract egg Remove egg

Sheep 1 cell nucleus

Therefore, the correct answer is A: the offspring would be genetically identical to sheep 2.
Let’s recap some of the key points we have covered in this explainer.

Key Points

Artificial reproduction is the creation of life by artificial means.

IVF is the process of combining eggs and sperm in a laboratory setting to create embryos that
can be transferred to the uterus in hopes of achieving a pregnancy.

Gametes such as eggs and sperm can be stored in gamete banks.

Artificial reproduction can also be used in farm animals such as cattle.
Somatic cell nuclear transfer is a technique used to clone animals.

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