Cell differentiation (1)

Stem cells become specialized through differential gene expression. Stem cells become
specialized because different genes in their DNA become active (are turned on)- in other
words, they express different genes to produce different proteins.

1
Review: Mendel's basic model

The basic principles of Gregor Mendel’s model of inheritance have held up for over a
century. They can explain how many different characteristics are inherited, in a wide range of
organisms including human beings.
Some of the key elements of Mendel’s original model were:

1. Heritable traits are determined by heritable factors, now called genes. Genes come in
pairs (that is, are present in two copies in an organism).
2. Genes come in different versions, now called alleles. When an organism has two
different alleles of a gene, one (the dominant allele) will hide the presence of the other
(the recessive allele) and determine appearance.
3. During gamete production, each egg or sperm cell receives just one of the two gene
copies present in the organism, and the copy allocated to each gamete is random (law
of segregation).
4. Genes for different traits are inherited independently of one another (law of
independent assortment).

These rules still form the foundation of our understanding of inheritance—that is, how traits
are passed on and how an organism's genotype (set of alleles) determines its phenotype
(observable features). However, we now know of some exceptions, extensions, and
variations, which must be added to the model in order to fully explain the inheritance patterns
we see around us.

An example of variations on Mendel’s rules involve single genes is multiple alleles.

 Multiple alleles: Mendel studied just two alleles of his pea genes, but real
populations often have multiple alleles of a given gene. Multiple Alleles are three or
more possible alleles for one individual trait.
 An excellent example of multiple allele inheritance is human blood type. Blood type
exists as four possible phenotypes: A, B, AB, & O. There are 3 alleles for the gene
that determines blood type. These alleles have different patterns of inheritance (both
A and B alleles are dominant to O which is recessive; Whereas A and B alleles are
codominant.
 Codominance. Two alleles may be simultaneously expressed when both are present,
producing proteins and in turn both contribute to determining the phenotype rather
than one fully determining the phenotype

2
The A, B and O alleles all produce a basic antigen on the surface of red blood cells

 The A and B alleles are co-dominant and each modify the structure of the antigen to
produce different variants
 The O allele is recessive and does not modify the basic antigenic structure
 When representing blood group alleles, the letter I is used to represent the different
antigenic forms

 A allele = IA ; B allele = IB ; O allele = i (recessive)

The genotypes for the different blood groups can be summarised as follows:

Polygenic traits

Polygenic inheritance refers to a single inherited phenotypic trait that is controlled by two or
more different genes. Polygenetic traits may display a range of possible phenotypes,
determined by a number of different genes and the interactions between them.
Height and other similar features (eye color, hair color, and intelligence) are controlled not
just by one gene, but rather, by multiple (often many) genes that each make a small
contribution to the overall outcome (a recent study found over 400 genes linked to variation
in height). In an additional complication, height doesn’t just depend on genetics: it also
depends on environmental factors, such as a child’s overall health and the type of nutrition he
or she gets while growing up.

Dihybrid (digenic) inheritance (two different genes that are inherited completely
independently of each other = unlinked)

3
According to the law of independent assortment, pairs of alleles are inherited independently
of one another if their gene loci are on separate chromosomes – these genes are said to be
unlinked.

 This is due to the random orientation of homologous pairs during metaphase I of

meiosis

The independent segregation of unlinked genes results in a greater number of potential

gamete combinations, as well as a greater variety of possible phenotypes

 This also results in more complex inheritance patterns (e.g. monohybrid versus
dihybrid crosses)

A dihybrid cross determines the genotypic and phenotypic combinations of offspring for two
particular genes that are unlinked.

 Because there are two genes, each with two alleles, there can be up to four different
gamete combinations

The easiest way to work out potential gamete combinations in a dihybrid cross is to use the
FOIL method:

 FOIL = First / Outside / Inside / Last

4
Calculation of the predicted genotypic and phenotypic ratio of offspring of dihybrid crosses
involving unlinked autosomal genes.

Example 1

5
Example 2

There are some occasions where the ratios of breeding experiments involving the inheritance
of two pairs of contrasting characteristics at the same time are not what you expect or
different from the theoretical ratios. Some reasons that explain this:

 Small sample size

 Experimental error
 Random process
 Genes are on the same chromosome (linked genes) rather than unlinked.

6
Gene linkage

When genes are close together on the same chromosome, they are said to be linked. That
means the alleles, or gene versions, already together on one chromosome will be inherited as
a unit more frequently than not

Facts:
 Linked genes will tend to be inherited together (don’t independently assort) and hence
don’t follow normal Mendelian inheritance for a dihybrid cross
 Instead the phenotypic ratio will be more closely aligned to a monohybrid cross as the
two genes are inherited as a single unit
 Linked genes may become separated via recombination (due to crossing over during
synapsis in meiosis I)

7
When genes are on the same chromosome but very far apart, they assort independently due to
crossing over (homologous recombination). Crossing over can put new alleles together in
combination on the same chromosome, causing them to go into the same gamete. When
genes are far apart, crossing over happens often enough that all types of gametes are
produced with 25% frequency.

When genes are very close together on the same chromosome, crossing over still occurs, but
the outcome (in terms of gamete types produced) is different. Instead of assorting
independently, the genes tend to "stick together" during meiosis. That is, the alleles of the
genes that are already together on a chromosome will tend to be passed as a unit to gametes.
In this case, the genes are linked. For example, two linked genes might behave like this:

8
Why are the recombinant gamete types rare?
The basic reason is that crossovers between two genes that are close together are not very
common. Crossovers during meiosis happen at more or less random positions along the
chromosome, so the frequency of crossovers between two genes depends on the distance
between them. A very short distance is, effectively, a very small "target" for crossover
events, meaning that few such events will take place (as compared to the number of events
between two further-apart genes).

Recombinants of linked genes are those combinations of genes not found in the parents

 Recombinants occur as a result of crossing over of genetic material during prophase I

of meiosis
 If linked genes become separated by a chiasma, there will be an exchange of alleles
between the non-sister chromatids
 This creates new allele combinations that are different to those of the parent

The frequency of recombinant phenotypes within a population will typically be lower than
that of non-recombinant phenotypes

 This is because crossing over is a random process and chiasmata do not form at the
same locations with every meiotic division

9
The relative frequency of recombinant phenotypes will be dependent on the distance between
linked genes

 Recombination frequency between two linked genes will be greater when the genes
are further apart on the chromosome
 This is because there are more possible locations where a chiasma could form
between the genes

10
 Interaction between genes and their environment (2)

 The phenotype is the observable characteristics or traits of an organism. It is

predominantly determined by the organism’s genotype (allele combination) for each
particular feature.
 However environmental factors may also influence the expression of characteristics

Example 1:

 Hydrangeas change colour depending on the pH of the soil (acidic soil = blue flower ;
alkaline soil = pink flower)
 Human skin colour is determined by the expression of melanin pigment, but levels
can change depending on sun exposure

Example 2 (LAC operon in E.Coli)

 The lac operon is an operon, or group of genes with a single promoter (transcribed as
a single mRNA). The genes in the operon encode proteins that allow the bacteria to
use lactose as an energy source.

 The lac operon of E. coli contains genes involved in lactose metabolism. It's
expressed only when lactose is present and glucose is absent.
 Two regulators turn the operon "on" and "off" in response to lactose and glucose
levels: the lac repressor and catabolite activator protein (CAP).
 The lac repressor acts as a lactose sensor. It normally blocks transcription of the
operon (in the absence of lactose), but stops acting as a repressor when lactose is
present.
 Catabolite activator protein (CAP) acts as a glucose sensor. It activates
transcription of the operon, but only when glucose levels are low.

E. coli bacteria can break down lactose, but it's not their favorite fuel. If glucose is around,
they would much rather use that. Glucose requires fewer steps and less energy to break down

11
than lactose. However, if lactose is the only sugar available, the E. coli will go right ahead
and use it as an energy source
To use lactose, the bacteria must express the lac operon genes, which encode key enzymes
for lactose uptake and metabolism. To be as efficient as possible, E. coli should express the
lac operon only when two conditions are met:

 Lactose is available, and

 Glucose is not available

The lac operon contains three genes: lacZ, lacY, and lacA. These genes are transcribed as a
single mRNA, under control of one promoter
In addition to the three genes, the lac operon also contains a number of regulatory DNA
sequences. These are regions of DNA to which particular regulatory proteins can bind,
controlling transcription of the operon

 The promoter is the binding site for RNA polymerase, the enzyme that performs
transcription.
 The operator is a negative regulatory site bound by the lac repressor protein. The
operator overlaps with the promoter, and when the lac repressor is bound, RNA
polymerase cannot bind to the promoter and start transcription.
 The CAP binding site is a positive regulatory site that is bound by catabolite
activator protein (CAP). When CAP is bound to this site, it promotes transcription by
helping RNA polymerase bind to the promoter

The lac repressor is a protein that represses (inhibits) transcription of the lac operon. It does
this by binding to the operator, which partially overlaps with the promoter. When bound, the
lac repressor gets in RNA polymerase's way and keeps it from transcribing the operon.

When lactose is not available, the lac repressor binds tightly to the operator, preventing
transcription by RNA polymerase. However, when lactose is present, the lac repressor loses
its ability to bind DNA. It floats off the operator, clearing the way for RNA polymerase to
transcribe the operon

12
 ↳

Example 3 (hair color in Himalayan rabbits or Siamese cats)

• To make melanin in animals, an enzyme called tyrosinase enzyme is required

• This enzyme catalyzes the conversion of tyrosine a.a into melanin (last stage in a multistep
reaction)
• If the alleles for tryosinase are mutated, tryosinase is produced but it is inactivated at room
temperature. In turn no melanin would be produced and they end up in having white fur.
• However, at the very tips of their body that are colder than elsewhere,tyrosinase is active
and melanin is produced making these areas darker than the rest of the body.

13
Example 4:

Jennifer and Karen are identical twins; Identical twins look exactly the same because each
twin shares the same genes as their identical sibling.
If both girls have genetic information (genes) associated with obesity, does this mean that
they will have no control over their weight? Or, if the girls are adopted by different families
and have different life experiences, will their environment dictate how much they will
ultimately weigh?

Genes can carry instructions that can make it more likely for you to develop certain illnesses
or conditions. For example, Jennifer and Karen both have genes associated with obesity.
Their genes could tell their body to:

 increase the size of their fat cells or dictate how they use fat in their body
 release chemicals (like hormones) which control hunger and appetite
 influence behavior as Jennifer and Karen interact with their environment. For
example, if Karen begins to gain weight, she may seek out fewer opportunities to
exercise because going to the gym makes her feel uncomfortable

Suppose Jennifer is raised by wealthy parents who have access to the best, healthiest foods.
Her parents cook nutritious meals like vegetable risotto and lentil soup and limit the amount
of sugar, salt, and fat their daughter consumes. Jennifer learns to love fruits and vegetables,
and doesn’t crave excessively salty or sweet foods. Her parents have a significant amount of
time to play with her and teach her to live an active lifestyle. By eating nutritious foods and
staying physically active, the genes increasing her chance of developing obesity are not
expressed and she never develops obesity.
Karen however is raised by low-income parents who live in an area where fresh, healthy food
is scarce and expensive. Because her parents can’t afford fruits and vegetables, Karen eats a
lot of frozen, packaged meals and fast food, which are higher in sugar, fat, and salt. Her
parents both work multiple jobs, and Karen spends her time alone in front of the television.
Karen’s eating and physical activity habits enhance the expression of her genes for obesity

14
The characteristics (physical traits and behaviors) that you are born with and what you
experience throughout your life are both important. Your characteristics can impact your
experiences and your experiences can impact your characteristics.

(Have a look at the twin studies example in your textbook page 200)

Variations

• Variation is the small differences that exist between individuals. Even members of the same
species have slightly different characteristics. Variation is partly due to genes and partly due
to differences in the environment. For example humans have the same general shape and the
same set of body organs, but some features differ from one person to another such as height,
weight, eye and hair color, shape of the nose, language, knowledge, and skills.
• Some of the variable features may be inherited and we call them inherited characteristics
such as the tendency to develop diseases and the permanent color of the skin.
• Some, however, such as the temporary suntan or scars can’t be inherited and we call them
acquired characteristics.
• Acquired characteristics can be changed whereas inherited characteristics can’t be usually
altered

Types of variation

Variation could be either discontinuous or continuous variation.

1) Discontinuous variation
 An organism either has the characteristic or doesn’t have it.
 There is no range of these characteristics between extremes(no intermediates). An
organism is placed into specific categories. There is no disagreement about the
categories.
 The characteristics are qualitative; they can’t be measured.
 The characteristics are the result of one gene only (monogenic); they are not affected
by the environment.
 Blood groups are a good example: you are either one blood group or another - you
can't be in between. Other examples include: gender (male or female), ability to roll
your tongue
 The bar chart that results cannot become a smooth curve because there are no
intermediate groups

15
2) Continuous variation
 Every organism within the population shows the characteristic but to a different
extent. The characteristic can have any value within a range (intermediates). Different
scientists might well disagree about which category any single organism falls into.
 Characteristics are quantitative; they can be measured
 These characteristics result from several acting genes (polygenic) or from both genes
and the environment.
 Height is an example of continuous variation: individuals can have a complete range
of heights, for example, 1.6, 1.61, 1.62, 1.625 etc metres high. Other examples of
continuous variation include: Weight; Hand span; Shoe size; nose size; skin color
(any shade from very dark to very pale)
 The bar chart that results can be redrawn as a smooth curve because there are possible
intermediate classes between the extremes. The curve is a bell shaped curve.
 When studying continuous variation in a population large sample size is taken
(chance can affect the result), randomized sample (for accuracy and to remove bias)

 When the characteristics are measured for any group of organisms, the highest
percentage of the population are intermediates, and lowest percentage is of low and
high extremes. We say that the characteristic follows normal distribution.

16
 Controlling gene expression (3)

Gene expression is when a gene in DNA is "turned on," that is, used to make the protein it
specifies. Not all the genes in your body are turned on at the same time, or in the same cells
or parts of the body
Eukaryotic gene expression involves many steps, and almost all of them can be regulated.
Different genes are regulated at different points, and it’s not uncommon for a gene
(particularly an important or powerful one) to be regulated at multiple steps.

 Chromatin accessibility. The structure of chromatin (DNA and its organizing

proteins) can be regulated. More open or “relaxed” chromatin makes a gene more
available for transcription.
 Transcription. Transcription is a key regulatory point for many genes. Sets of
transcription factor proteins bind to specific DNA sequences in or near a gene and
promote or repress its transcription into an RNA.
 RNA processing. Splicing to an RNA molecule can be regulated before it can exit
from the nucleus. Different mRNAs may be made from the same pre-mRNA by
alternative splicing
 RNA stability. The lifetime of an mRNA molecule in the cytosol affects how many
proteins can be made from it. Small regulatory RNAs called miRNAs can bind to
target mRNAs and cause them to be chopped up.
 Translation. Translation of an mRNA may be increased or inhibited by regulators.
For instance, miRNAs sometimes block translation of their target mRNAs (rather than
causing them to be chopped up).
 Protein activity. Proteins can undergo a variety of modifications, such as being
chopped up or tagged with chemical groups. These modifications can be regulated
and may affect the activity or behavior of the protein.

Although all stages of gene expression can be regulated, the main control point for many
genes is transcription.

1- Transcription

Transcription is the process where a gene's DNA sequence is copied (transcribed) into an
RNA molecule. Transcription is a key step in using information from a gene to make a
protein

During transcription of a gene, the enzyme RNA polymerase, which makes a new RNA
molecule from a DNA template, must attach to the DNA of the gene. It attaches at a spot
called the promoter.
In humans and other eukaryotes, RNA polymerase can attach to the promoter only with the
help of proteins called basal (general) transcription factors. They are part of the cell's core
transcription toolkit, needed for the transcription of any gene.

17
What are transcription factors?

Transcription factors are proteins that regulate the transcription of genes—that is, their
copying into RNA, on the way to making a protein; in other words, transcription factors help
turn specific genes "on" or "off" by binding to nearby DNA.

Some transcription factors are activators and in turn boost a gene's transcription. Others are
repressors and in turn decrease transcription.

Groups of transcription factor binding sites called enhancers and silencers can turn a gene
on/off in specific parts of the body

The binding sites for transcription factors are often close to a gene's promoter. However, they
can also be found in other parts of the DNA, sometimes very far away from the promoter, and
still affect transcription of the gene.

Activators
Some transcription factors activate transcription. For instance, they may help the general
transcription factors and RNA polymerase bind to the promoter, as shown in the diagram
below

Repressors
Other transcription factors repress transcription. This repression can work in a variety of
ways. As one example, a repressor may get in the way of the basal transcription factors or
RNA polymerase, so they can't bind to the promoter or begin transcription.

18
Transcription factors also regulate gene expression by changing the structure of chromatin ,
making it less or more open to RNA polymerase. For instance, how tightly the DNA of the
gene is wound around its supporting proteins to form chromatin can affect a gene's
availability for transcription.

Several transcription factors may be involved in the control of a single gene. Sometimes a
single transcription factor may control the activity of several genes. It may stimulate the
expression of one gene and suppress the expression of another. This is a way in control of
multiple genes is brought about. It means each gene can be switched on or off at different
stages of the development of an organism, in different cell types and under different
circumstances in the body.

2- RNA splicing

 When a eukaryotic gene is transcribed in the nucleus, the primary transcript (freshly
made RNA molecule) isn't yet considered a messenger RNA. Instead, it's an
"immature" molecule called a pre-mRNA.
 The pre-mRNA has to go through some modifications to become a mature mRNA
molecule that can leave the nucleus and be translated. These include splicing.
 Most pre-mRNA molecules have sections that are removed from the molecule, called
introns (non-coding DNA sequences), and sections that are linked or together to
make the final mRNA, called exons (coding DNA sequences). This process is called
splicing
 Introns are recognized and removed by a protein-and-RNA complex called the
spliceosome. Introns can be viewed as "junk" sequences that must be cut; The exons
are pasted together by the spliceosome to make the final, mature mRNA that is
shipped out of the nucleus

 RNA splicing does allow for a process called alternative splicing, in which more
than one mRNA can be made from the same gene. Through alternative splicing, we
(and other eukaryotes) can sneakily encode more different proteins than we have
genes in our DNA.

 In the process of alternative splicing, different portions of an mRNA can be selected

for use as exons. This allows either of two (or more) mRNA molecules to be made
from one pre-mRNA.

19
  Alternative splicing is not a random process. Instead, it's typically controlled by
regulatory proteins. The proteins bind to specific sites on the pre-mRNA and "tell" the
splicing factors which exons should be used. Different cell types may express
different regulatory proteins, so different exon combinations can be used in each cell
type, leading to the production of different proteins.

3-Post-translation control

There are also regulatory mechanisms that act on proteins that have already been made. In
these cases, an "edit" to the protein – such as removal of amino acids, or addition of a
chemical modification – can lead to a change in its activity or behavior. These processing and
modification steps can be targets for regulation

Epigenetics

Your genes play an important role in your health, but so do your behaviors and environment,
such as what you eat and how physically active you are. Epigenetics is the study of how your
behaviors and environment can cause changes that affect the way your genes work. Unlike
genetic changes, epigenetic changes are reversible and do not change your DNA sequence,
but they can change how your body reads a DNA sequence.

Gene expression refers to how often or when proteins are created from the instructions within
your genes. While genetic changes can alter which protein is made, epigenetic changes affect
gene expression to turn genes “on” and “off.” Since your environment and behaviors, such as
diet and exercise, can result in epigenetic changes, it is easy to see the connection between
your genes and your behaviors and environment.

Types of DNA

On a different note, when it comes to gene expression and DNA, you can basically think of
DNA as coming in two flavors:
1- densely packed, and transcriptionally inactive DNA, which is called
heterochromatin.
2- less dense, transcriptionally active DNA, which is euchromatin.

20
How Does Epigenetics Work?

Epigenetic changes affect gene expression in different ways. Types of epigenetic changes
include:

1. DNA Methylation
 DNA methylation involves the addition of a methyl group, which is a carbon with
three hydrogens, to the cytosine, DNA nucleotides, by an enzyme appropriately called
methyltransferase.
 This typically occurs in cytosine-rich sequences called CpG islands. Don't forget that
cytosine pairs with g, guanine, so that's why they're cg islands that you'll find.
 DNA methylation stably alters the expression of genes (blocks the transcriptional
proteins that attach to DNA to “read” the gene and modifies the histones,
forming condensed, inactive heterochromatin that is basically transcriptionally
silent)
 It occurs as cells divide and differentiate from embryonic stem cells into specific
tissues. And so this is essential for normal development, and is associated with other
processes, such as genomic imprinting, and x-chromosome inactivation.
 Abnormal DNA methylation has been implicated in carcinogenesis, or the
development of cancer, so you can see how the normal functioning of DNA
methylation is a critical regulatory mechanism for our cells.
 This chemical group can be removed through a process called demethylation.
Typically, methylation turns genes “off” and demethylation turns genes “on.”

2. Histone modification

DNA wraps around proteins called histones (positively charged) to form chromatin..
DNA wrapped tightly around histones i.e supercoiled and condensed
(heterochromatin) cannot be accessed by proteins that “read” the gene. Some genes
are wrapped around histones and are turned “off” while some genes are not wrapped
around histones i.e are loosely packed (active chromatin=euchromatin) and are turned
“on.”

a. Histone acetylation: is the addition of an acetyl group; acetylation occurs at the

amino terminal tails of these histone proteins (lysine amino acid) by an enzyme called
histone acetyltransferase. And this is a reversible modification, so the acetylation of
histones is sort of kept in balance by another enzyme that removes these acetyl
groups, which is called histone deacetylase. The acetylation of histones leads to
uncoiling of this chromatin structure, and this allows it be accessed by transcriptional
machinery for the expression of genes.
b. Histone methylation: is a process by which methyl groups are transferred to amino
acids of histone proteins that make up nucleosomes. Methylation of histones can
either increase or decrease transcription of genes, depending on which amino acids in
the histones are methylated, and how many methyl groups are attached. Methylation
events that weaken chemical attractions between histone tails and DNA increase
transcription because they enable the DNA to uncoil from nucleosomes so that
transcription factor proteins and RNA polymerase can access the DNA. This process
is critical for the regulation of gene expression that allows different cells to express
different genes.

21
 3. Non-coding RNA (ncRNA)

 Your DNA is used as instructions for making coding and non-coding RNA. Coding
RNA is used to make proteins. Non-coding RNA helps control gene expression by
attaching to coding RNA, along with certain proteins, to break down the coding RNA
so that it cannot be used to make proteins. Non-coding RNA may also recruit proteins
to modify histones to turn genes “on” or “off.
 X-chromosome inactivation occurs randomly for one of the two X chromosomes in
female cells during development. Inactivation occurs when ncRNA called X-inactive
specific transcript (Xist) transcribed from the Xist gene on the X chromosome from
which it is expressed spreads to coat the whole X chromosome. The chromosome
supercoils, condenses, and changes to inactive Barr body.
 X-inactivation ensures that females, like males, have one functional copy of the X
chromosome in each body cell. Because X-inactivation is random, in normal females
the X chromosome inherited from the mother is active in some cells, and the X
chromosome inherited from the father is active in other cells

22
 Stem cells and their uses(4 and 5)

Early stages of development

 After fertilization, a complex series of events that will eventually lead to the birth of
a fully formed new individual
 In humans, the fertilized egg or the zygote has the potential to develop into the 216
different cell types needed for the entire new person
 The first stage of the process is called cleavage (special kind of mitosis) resulting in a
small ball of identical and undifferentiated cells called a blastocyst
 The tiny cells of the embryo are known as stem cells.

Stem cells

 Stem cells are unspecialized cells that can develop into any other cell type.
 Stem cells divide by mitosis to become new cells which then become specialized.
 All multicellular organisms have some form of stem cells
 There are different types of stem cells:

1. Totipotent is a cell that is able to divide and produce all of the differentiated cells in
an organism (including all specialized cells in an organism and the extrasmbryonic
cells that form the placenta and the umbilical cord).
Spores and Zygotes are examples of totipotent cells.
In the spectrum of cell potency, totipotency represents the cell with the greatest
differentiation potential.

2. Pluripotent is a cell that is able to differentiate into any of the three germ layers:
endoderm (interior stomach lining, gastrointestinal tract, the lungs), mesoderm
(muscle, bone, blood, urogenital), or ectoderm (epidermal tissues and nervous
system) or in other words it is the ability of a stem cell to produce all the specialized
cells in an organism but not the extraembryonic cells.

3. Multipotent is a cell that develops into limited cell types; they form one type of
tissue cells

Note

 The process by a which a cell becomes specialized is called differentiation

 De-differentiation is the formation of simple undifferentiated cells from specialized
cells
 The ability of stem cells to differentiate into specialized cells is called potency

23
  Cells of the inner mass are pluripotent; they can form all cell types of the human
body except the placenta; they are the source of embryonic stem cells
 The outer cell layer (trophoblast) forms the placenta

24
Uses of stem cells

1. In production of specific tissues and organs that could be used in transplants

2. Injected to specialized tissues or organs to replace damaged tissues or cells
3. Used for scientific research to study how cell differentiation occurs, help to
understand how genetic disorders develop so that a strategy of prevention and
treatment could be worked out.
4. Provide differentiated cells to test new drugs in the laboratory, rather than testing
drugs on animals

Sources or types of human stem cells

There are three potential sources of human stem cells: adult tissue, umbilical cord blood,
and embryonic stem cells

A. Adult stem cells

1. They are obtained from the body tissues of an adult (bone marrow, fat tissue, brain,
skin, hair follicles)
2. They can be obtained in a relatively simple operation-with very little risk involved,
but quite a lot of discomfort. The donor is anaesthetized, a needle is inserted into
the center of a bone (usually the hip) and a small quantity of the bone marrow is
removed
3. Are found in tissues to replace damaged or dead cells and in regeneration and
wound healing

Advantages
 No ethical issues regarding their harvesting and use
 They seem to be abundant in one single human as the wound healing process
continues to occur throughout life
 If used in treatment, there seems to be no or little problems of rejection

Disadvantages
 Adult stem cells aren’t as flexible as embryonic stem cells-they can only
develop into a limited range of cells
 In 2006, scientists were able to reprogramme specialized skin cells into
pluripotent stem cells; these cells are called induced pluripotent stem cells
(iPSCs) and are good alternatives to embryonic stem cells in research;
however, there is an evidence that they increase the risk of tumours
 Are so hard to identify and isolate from other cells in the tissues.

25
 B. Embryonic stem cells

1. These are obtained from early embryos

2. Once the embryos are approximately 4 to 5 days old, stem cells are removed from
them and the rest of the embryo is destroyed.

Advantages
 Embryonic stem cells can develop into all types of specialized cells (totipotent
or pluripotent);
 It is easier to identify and isolate embryonic stem cells

Disadvantages
 After harvesting embryonic stem cells, the rest of the embryo is discarded;
this raises controversy against the process as this is mirrored to abortion and
considered to be unethical
 Limited supply of spare embryos to extract stem cells
 Problems associated with tissue rejection or tumour development

Sources of embryonic stem cells

 Spare embryos created by in vitro fertilization (IVF) for fertility treatment
(used in scientific research into serious diseases and for understanding the
processes of differentiation and de-differentiation)
 Therapeutic cloning is useful to overcome tissue rejection
 Animal human embryos used in understanding differentiation and de-
differentiation

26
 C. Umbilical cord blood stem cells

1. Harvested from the cord blood of umbilical cord and placenta soon after
birth
2. Stem cells are pluripotent; they are mainly hematopoetic stem cells (like
adult bone marrow stem cells) meaning that they can make different types of
blood cells.
3. Used to treat the same baby later in life

Advantages
 Have a higher potency than bone marrow or other adult stem cells
 Low chance of tissue rejection
 There is a ready supply of cord blood, and collection of it doesn’t
harm the mother or the baby.

Disadvantages
 Limited source of cord blood stem cells
 The cord blood stem cells take a longer time to graft into the patient
than do bone marrow transplants
 Collecting and storing of cord blood is expensive
Risks of stem cell use

1. Stem cells has an increased chance to become cancerous

2. Rejection by the recipient
3. Possible route to infection

Stem cells could be used to treat some diseases (read textbook page 214)

1. Stem cells can develop into any specialized cell type, so scientists think they could be
used to replace damaged tissues in a range of diseases.
2. Some stem cell therapies already exist. For example, the treatment of leukemia (a
cancer of the bone marrow) kills all the stem cells in the bone marrow, so bone
marrow transplants can be given to patients to replace them.
3. Scientists are researching the use of stem cells as a treatment for lots of conditions,
including:
 Spinal cord injuries-stem cells could be used to repair damaged nerve tissue
 Heart disease and damage caused by heart attacks-stem cells could be used to
replace damaged heart tissue
1. People who make decisions about the use of stem cells in medicine and research
have to consider the potential benefits of stem cell therapies:
 They could save many lives-e.g. many people waiting for organ transplants die
before a donor organ becomes available. Stem cells could be used to grow organs
for people awaiting transplants
 They could improve the quality of life for many people-e.g. stem cells could be used
to replace damaged cells in the eyes of people who are blind

27
Stem cell cloning (therapeutic cloning)
Therapeutic cloning is an experimental technique to produce healthy tissue using stem cells.
The aim is to treat someone with a disease caused by faulty cells such as type I diabetes.

Steps:

1. A scientist extracts the nucleus from an egg

2. The nucleus holds the genetic material for a human or laboratory animal
3. The scientist then takes a somatic cell, which is any body cell other than an egg or
sperm, and also extract the nucleus from this cell
In practical human applications, the somatic cell would be taken from a patient who
requires a Stem Cell Transplant to treat a health condition or disease.
4. The nucleus that is extracted from the somatic cell in the patient is then inserted
into the egg, which had its nucleus previously removed
5. In a very basic sense, it's a procedure of substitution. The egg now contains the
patient's genetic material, or instructions
6. It is stimulated to divide and shortly thereafter forms a cluster of cells known as a
blastocyst

28
29
Benefits of therapeutic cloning

 The cells removed are pluripotent. Pluripotent Cells, provided with the appropriate
signal, can give rise to all cells in the body with the exception of the embryo.
 Another distinct advantage to this type of therapy is that the risk of immunological
rejection is alleviated because the patient's own genetic material is used. If a cell
line were created with cells from another individual, the patient's body would be
more likely to recognise the foreign proteins and then wage an attack on the
transplanted cells. The ultimate consequence would be a rejected stem cell
transplant. This is one of the major challenges of organ transplants, alongside the
fact that there is a huge shortage of available organs for those who require the
procedure. This means that therapeutic cloning has the potential to dramatically
reduce the wait times for organ transplants as well as the immunological concerns
associated with organ transplant therapy.
 Used in scientific research (details mentioned before in uses of stem cells)

Problems with therapeutic cloning

 One problem with therapeutic cloning is that many attempts are often required to
create a viable egg. The stability of the egg with the infused somatic nucleus is poor
and it can require hundreds of attempts before success is attained.
 Therapeutic cloning does result in the destruction of an embryo after stem cells are
extracted and this destruction has stirred controversy over the morality of the
procedure. Some argue that the pros outweigh the cons with regards to treating
disease whilst others have likened the destruction to an abortion. Still others state
that this doesn't change the fact the embryo could potentially be a human being and
so destruction of the embryo is no different than destruction of a human life.
 Because reproductive cloning does utilise therapeutic cloning as the primary step,
there is also still fear that given our knowledge base to perform reproductive
cloning, a scientist may attempt to move beyond therapeutic cloning to creation of a
human being.
 In certain cases, adult stem cells are clearly preferable because of the risk that the
use of volatile embryonic stem cells will cause tumors of various types.

30
Ethics of stem cells (read pages 214 and 215 of your textbook)

Pros
1. Many people think that it is a major breakthrough in science with the potential to
change the healthcare.
2. The vast majority of human embryos never make it beyond the early stages of
development to form living babies, so the argument for using a small number of
early embryos is acceptable in this context.
3. Adult stem cells do not offer a good alternative because they are much limited in
their scope for forming new and different tissues

Cons
1. Other people feel that the use of embryos is wrong and an abuse of human rights
2. Many religious groups think it is wrong to use a potential human in this way
3. Some others feel that every early embryo has the potential to become a living
human being and should be afforded the same human rights as a fully grown adult.
4. Others have strong religious convictions that using embryos is killing and therefore
wrong. They think that no medical advances are worth the moral evil of using
embryonic tissue as a source of stem cells.
5. Many people feel that the use of adult stem cells offers an exciting and acceptable
possible alternative to embryonic cells and they campaign for research funding to be
directed to projects using these ethically less sensitive cells.
6. The use of embryonic stem cells from the umbilical cord of newly born babies may
help to overcome many of these reservations

Society makes decisions about the use of stem cells

1. Embryonic stem cells could be really useful in medicine, but research into their use
raises many ethical issues
2. Society has to consider all the arguments for and against stem cell research before
allowing it to go ahead
3. To help society make these decisions, regulatory authorities have been
established to consider the benefits and ethical issues surrounding embryonic stem
cell research
4. The work of regulatory authorities includes:
 Looking at proposals of research to decide if they should be allowed-this ensures
that any research involving embryos is carried out for a good reason (fertility
treatment or scientific research into human illnesses). This also makes sure
research isn’t unnecessarily repeated by different groups
 Licensing and monitoring centers involved in embryonic stem research-this ensures
that only fully trained staff carry out the research. These staff will understand the
implications of the research and won’t waste precious resource, such as embryos.
This also helps to avoid unregulated research
 Producing guidelines and codes of practice-this ensures all scientists are working in
a similar manner (if scientists don’t use similar methods their results can’t be
compared). It also ensures methods of extraction are controlled.

31
  Monitoring developments in scientific research and advances- this ensures that any
changes in the field are regulated appropriately and that all the guidelines are up to
date with the latest in scientific understanding.
 Providing information and advice to governments and professionals- this helps to
promote the science involved in embryo research, and it helps society to
understand what’s involved and why its important.
 The written consent of the donors of the embryos must also be given and this must
clearly show that they have consented to the uses to which their embryos will be put
 No embryo can be kept or used after 14 days of its development. This is because the
nervous system does not start to develop till then.

32