Stem Cells

Stem Cells
Dr. Surya Prakash G. Ponnam,

M.Sc., Ph.D.
Assistant Professor
Department of Molecular Biology and
Biotechnology,
Tezpur University-Napaam
Sonitpur, Assam, India-78028
1
Introduction
Cells that can differentiate into other types of cells and can
divide to produce more of the same type of stem cells.
They are found in multicellular organisms.
Research into stem cells grew out of findings by Ernest A.
McCulloch and James E. Till at the University of Toronto in the
1960s.
The classical definition of a stem cell requires that it possesses
two properties
a) Self-renewal: the ability to go through numerous cycles of
cell division while maintaining the undifferentiated state.
b) Potency: the capacity to differentiate into specialized cell
types.
Self-renewal
Two major mechanisms exist to ensure that a stem cell
population is maintained:
1.Obligatory asymmetric replication
A stem cell divides into one mother cell that is identical to the
original stem cell, and another daughter cell that is
differentiated.
2. Stochastic differentiation
When one stem cell develops into two differentiated daughter
cells, another stem cell undergoes mitosis and produces two
stem cells identical to the original.
Potency
The potential to differentiate into different cell types of the stem
cell
Totipotent (omnipotent)
stem cells can differentiate into embryonic and extraembryonic cell
types.
Such cells can construct a complete, viable organism.
These cells are produced from the fusion of an egg and sperm cell.
Cells produced by the first few divisions of the fertilized egg are
also totipotent.
Pluripotent stem cells
Descendants of totipotent cells and can differentiate into nearly all
cells, i.e. cells derived from any of the three germ layers.
Potency
Multipotent stem cells
can differentiate into several cell types, but only those of a
closely related family of cells.
Oligopotent stem cells
can differentiate into only a few cell types, such as lymphoid
or myeloid stem cells.
Unipotent cells
can produce only one cell type, their own, but have the
property of self-renewal, which distinguishes them from non-
stem cells (e.g. progenitor cells, which cannot self-renew
Identification of Stem cells
1. Stem cells are identified by their ability to regenerate
tissue.
2. Properties of stem cells can be illustrated in vitro, using
methods such as clonogenic assays, in which single cells
are assessed for their ability to differentiate and self-renew.
3. Stem cells can also be isolated by their possession of a
distinctive set of markers.
Embryonic stem cells
Embryonic stem cells (ESCs) are the cells of the inner cell mass of a
blastocyst, an early-stage embryo.
Human embryos reach the blastocyst stage 4–5 days post fertilization,
at which time they consist of 50–150 cells.
ESCs are pluripotent and give rise during development to all derivatives
of the three primary germ layers: ectoderm, endoderm and mesoderm.
 ESC can develop into each of the more than 200 cell types of the adult
body when given sufficient and necessary stimulation for a specific cell
type.
They do not contribute to the extra-embryonic membranes or the
placenta.
Mouse ES Cells
They are grown on a layer of gelatin as an extracellular matrix
and require the presence of leukemia inhibitory factor (LIF) in
serum media.
A drug cocktail containing inhibitors to GSK3B and the
MAPK/ERK pathway, called 2i, has also been shown to
maintain pluripotency in stem cell culture.
Human ES Cells
They are grown on a feeder layer of mouse embryonic fibroblasts and
require the presence of basic fibroblast growth factor (bFGF or FGF-2).
A hES is defined by the expression of several transcription factors and

cell surface proteins.
The transcription factors Oct-4, Nanog, and Sox2 form the core
regulatory network that ensures the suppression of genes that lead to
differentiation and the maintenance of pluripotency.
The cell surface antigens most used to identify hES cells are the
glycolipids stage specific embryonic antigen 3and 4 and the keratan
sulfate antigens Tra-1-60 and Tra-1-81.
Human ES Cells
 By using human embryonic stem cells to produce specialized cells like
nerve cells or heart cells in the lab, scientists can gain access to adult
human cells without taking tissue from patients. They can then study
these specialized adult cells in detail to try and catch complications of
diseases, or to study cells reactions to potentially new drugs.
 ES cells, being pluripotent cells, require specific signals for correct

differentiation—if injected directly into another body, ES cells will
differentiate into many different types of cells, causing a teratoma.
 Due to ethical considerations, many nations currently

have limitations/banned on either human ES cell research or the
Fetal embryonic stem cells
The primitive stem cells located in the organs of fetuses are referred
to as fetal stem cells.
There are two types of fetal stem cells:
Fetal proper stem cells
come from the tissue of the fetus proper and are generally obtained
after an abortion.
These stem cells are not immortal but have a high level of division
and are multipotent.
Extraembryonic fetal stem
cells come from extraembryonic membranes and are generally not
distinguished from adult stem cells.
These stem cells are acquired after birth, they are not immortal but
have a high level of cell division and are pluripotent.
Adult Stem cells
Also called somatic stem cells.
These are stem cells which maintain and repair the tissue in which they are
found.
They can be found in children, as well as adults.
Most adult stem cells are lineage-restricted (multipotent) and are generally
referred to by their tissue origin (mesenchymal stem cell, adipose-derived
stem cell, endothelial stem cell, dental pulp stem cell, etc.)
When subjected to single cell suspension culture, the cells will generate
clusters that are like embryoid bodies in morphology as well as gene
expression, including canonical pluripotency markers Oct4, Sox2,
and Nanog.
Pluripotent adult stem cells are rare and generally low in number, but they can
be found in umbilical cord blood and other tissues.
Bone marrow is a rich source of adult stem cells, which have been used in
treating several conditions including liver cirrhosis, chronic limb ischemia, etc.
The quantity of bone marrow stem cells declines with age and is greater
in males than females during reproductive years.
Most of the adult stem cell research to date has aimed to characterize
their potency and self-renewal capabilities.
Muse cells (multi-lineage differentiating stress enduring cells) are a
recently discovered pluripotent stem cell type found in multiple adult
tissues, including adipose, dermal fibroblasts, and bone marrow. While
rare, muse cells are identifiable by their expression of SSEA-3, a marker
for undifferentiated stem cells, and general mesenchymal stem cells
markers such as CD105.
Adult stem cell treatments have been successfully used for many years
to treat leukemia and related bone/blood cancers through bone marrow
transplants.
Adult stem cells are also used in veterinary medicine to treat tendon and
ligament injuries in horses.
Amniotic stem cells
Multipotent stem cells are also found in amniotic fluid.
These stem cells are very active, expand extensively without feeders
and are not tumorigenic. Amniotic stem cells are multipotent and can
differentiate in cells of adipogenic, osteogenic, myogenic,
endothelial, hepatic and neuronal lines.
Use of stem cells from amniotic fluid overcomes the ethical

objections to using human embryos as a source of cells.
Induced pluripotent stem cells (iPSC)
Adult stem cells have limitations with their potency
Reprogramming allows for the creation of pluripotent cells, induced

pluripotent stem cells(iPSCs), from adult cells. These are not adult stem
cells, but adult cells reprogrammed to give rise to cells with pluripotent
capabilities.
Using genetic reprogramming with protein transcription factors,

pluripotent stem cells with ESC-like capabilities have been derived.
The first demonstration of induced pluripotent stem cells was

conducted by Shinya Yamanaka et al., Kyoto University. They used the
transcription factors Oct3/4, Sox2, c-Myc, and Klf4 to reprogram mouse
Few Similarities/differences of IPSC
from ESC
Similarities
Pluripotency and differentiation potential
 the expression of pluripotency genes,
epigenetic patterns, embryoid body and teratoma formation,
and viable chimera formation
Differences
The chromatin of iPSCs appears to be more "closed" or methylated than that
of ESCs.
Similarly, the gene expression pattern between ESCs and iPSCs, or even iPSCs
sourced from different origins.
 There are thus questions about the "completeness" of re-programming and
the somatic memory of induced pluripotent stem cells. Despite this,
inducing adult cells to be pluripotent appears to be viable.
Applications of iPSCs
Like ESCs, they are pluripotent.
They thus have great differentiation potential; theoretically, they could
produce any cell within the human body (if reprogramming to
pluripotency was "complete").
Moreover, unlike ESCs, they potentially could allow doctors to create a
pluripotent stem cell line for each individual patient.
Frozen blood samples can be used as a valuable source of induced
pluripotent stem cells.
Patient specific stem cells allow for the screening for side effects before
drug treatment, as well as the reduced risk of transplantation rejection.
iPSCs hold create potential for future use in medical treatment and
research.
Acknowledgments
 Dept. of MB&BT, TU.
 All the original content creators whose images/

videos / text or any other information that has been
used in this presentation to make it effective.
Arise awake and stop not till the goal is reached
-----Swami Vivekananda
Thank You for your kind

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Stem Cells

Dr. Surya Prakash G. Ponnam,

A hES is defined by the expression of several transcription factors and

 ES cells, being pluripotent cells, require specific signals for correct

 Due to ethical considerations, many nations currently

Multipotent stem cells are also found in amniotic fluid.

Use of stem cells from amniotic fluid overcomes the ethical

Reprogramming allows for the creation of pluripotent cells, induced

Using genetic reprogramming with protein transcription factors,

The first demonstration of induced pluripotent stem cells was

 Dept. of MB&BT, TU.

 All the original content creators whose images/

Thank You for your kind

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