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Stem cell technology

Presentation · November 2017

DOI: 10.13140/RG.2.2.16332.92802

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 STEM CELL TECHNOLOGY---
A WORLD OF POSSIBILITIES

Department of Zoology
SAWAR KHAN Hazara University, Mansehra,
Pakistan
Email: sawarkhanzoo@gmail.com
 Some basic terms in stem cell biology
Cell differentiation
The allotment of functions to the cells. Cells having the clear cut identities are
said as differentiated. e.g, Neurons, skin cells and muscle cells etc.
Fate of cell
It describes what a cell will become in the course of normal development
Cell Potency (Latin; Potens= having power)
It is the developmental potential of a cell i.e, the ability of a cell to differentiate
into other cell types. The more cell types a cell can differentiate into the greater
its potency. It is also described as the gene activation potential with in a cell. Cell
potency may be of following types:
i) Totipotency (Latin; Totus=Entirely)
The ability of a single cell to divide and produce all of the cells in an organism.
e.g, zygote and spores
 Some basic terms in stem cell biology
ii) Pluripotency (Latin; Plurimus=very, many)
It refers to the cells having potential to differentiate into any of the three germ layers and
their derivatives. These cell are capable to differentiate into all types of body tissues except
extra embryonic tissues. Examples are the cells of inner cell mass.
iii) Multipotency
It refers to progenitor cells having potential to differentiate into multiple but limited cell
types. e.g, multipotent blood stem cells that can differentiate into various types of blood
cells.
iv) Oligopotency
It refers to the ability of progenitor cells to differentiate into a few cell types. e.g, lymphoid
cell can give rise to B and T cells only but not all the blood cells.
v) Unipotency
It refers to the capacity of cells to differentiate into only one cell type. e.g, hepatoblasts give
rise hepatocytes and osteoblasts give rise to osteocytes
 Stem Cells
 Cells that can give rise to the cells like
themselves or other cell types
 ‘Blank cells’ (unspecialized)
 Capable of dividing and renewing
themselves for long periods of time
(proliferation and renewal)
 Have the potential to give rise to
specialized cell types (differentiation)

11/13/2017
 Stem Cell Types
• Embryonic – ICM pluripotent: can form
almost any cell type in the human body

• Tissue-Specific (Adult) –
multipotent: can form only limited
types of cells (blood, brain, liver, etc.)

• Induced Pluripotent –generated by

reprogramming differentiated cells
•engineered by scientists to act like embryonic
stem cells
All Stem Cells Can…
1. Self-Renew

2. Differentiate
Self-Renewal (symmetric cell division)
Differentiation (asymmetric cell division)

Progenitor cell

Stem cell Stem cell

 Types of Stem cells based on Potency
• Totipotent stem cells---- Each cell in the 8-cell embryo can generate
every cell in the embryo as well as the extra-embryonic tissues

• Pluripotent stem cells--- The embryonic stem cells which can

generate every cell in the body except extra-embryonic tissues
• --from embryos that are 5-14 days old
• Multipotent stem cells ---- tissue-specific stem cells (Adult)
• Unipotent stem cells----only generate one cell type
 Pluripotency markers
• Oct3/4, Sox2, Nanog – transcription factors
involved with maintaining pluripotency
The original question--genome equivalence
Do all cells in the body have the same sets of
genes? OR
The genome of cells undergoes any stable
changes in the course of cell differentiation

Weissmann (1892)…. as cells embark on a

defined developmental pathway, they lose
genes no longer needed to be expressed
Spemann (1928)…. at least up to 8 cell stage,
nuclei are totipotent.
Somatic Cell Nuclear Transfer (SCNT)
• Developed the techniques
in 1950s
• (1) a method for enucleating
host eggs without destroying
them;
• (2) a method for isolating
intact donor nuclei; and
• (3) a method for transferring
such nuclei into the host egg
without damaging either the
nucleus or the oocyte.
• 30% of transplanted
blastula nuclei yielded
apparently normal
tadpoles,
• first to provide a clear test
of the genome
equivalence in
development
Conception in a Dish
In Vitro Fertilization
 In Vitro Fertilization

Day 1
 In Vitro Development

Day 1
 In Vitro Development

Day 2
 In Vitro Development

Day 2
 In Vitro Development

Day 3
 In Vitro Development

Day 4
 In Vitro Development

Day 5
Embryonic
Stem
Cells
 Blastocyst Diagram

11/13/2017 Dr. Hariom Yadav

Princeton University
 Reprogramming
• SCNT – somatic cell nuclear transfer
(reproductive cloning)
• iPS – induced pluripotent stem cells
Life is not a one-way journey
• Life there and back again

Sir John B. Gurdon

Gurdon, 1962 vs Briggs & King, 1952
Dolly the sheep

• In 1997, Ian Wilmut

announced that a
sheep had been
cloned from a
somatic cell nucleus
from an adult
female sheep.
 Reprogramming Firsts: SCNT
Frog:
Gurdon JB. (1962) Adult frogs derived from the nuclei of single
somatic cells. Dev Biol. 4:256-73.

Sheep:
Campbell KH, McWhir J, Ritchie WA, Wilmut I. (1996) Sheep
cloned by nuclear transfer from a cultured cell line. Nature.
380:64-6.

Human:
Tachibana M, et al. (2013) Human embryonic stem cells derived
by somatic cell nuclear transfer. Cell. 153:1228-38.
 First Isolation of ES cells
Mouse:
Evans MJ, Kaufman MH. (1981) Establishment in culture of pluripotential
cells from mouse embryos. Nature. 292:154-6.

Martin GR. (1981) Isolation of a pluripotent cell line from early mouse
embryos cultured in medium conditioned by teratocarcinoma stem cells.
P.N.A.S. U S A. 78:7634-8.

Human:
Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ,
Marshall VS, Jones JM. (1998) Embryonic stem cell lines derived from
human blastocysts. Science. 282:1145-7.
Dr. Shinya Yamanaka
 iPSC Functional confirmation

-and I did it…

 Reprogramming Firsts: iPS cells
Mouse:
Takahashi K, Yamanaka S. (2006) Induction of pluripotent stem
cells from mouse embryonic and adult fibroblast cultures by
defined factors. Cell. 126:663-76

Human:
Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda
K, Yamanaka S. (2007) Induction of pluripotent stem cells from
adult human fibroblasts by defined factors. Cell. 131:861-72.

Yu J, Vodyanik MA, Smuga-Otto K, Antosiewicz-Bourget J, Frane

JL, Tian S, Nie J, Jonsdottir GA, Ruotti V, Stewart R, Slukvin II,
Thomson JA. (2007) Induced pluripotent stem cell lines derived
from human somatic cells. Science. 318:1917-20.
 Generating iPS cells
• Express transcription factors:
Oct3/4, Sox2, Klf4 and c-Myc OR
Oct3/4, Sox2, Nanog and Lin28
• Initial de-differentiation and proliferation
(day 1-3, enhanced by Myc); histone modification
and chromatin reorganization
• 2nd wave of gene expression - stem cell and
development related genes (day 9-12); DNA
demethylation and X reactivation
 Pros and Cons to iPS cell
technology
• Pros:
– Cells would be genetically identical to patient or
donor of skin cells (no immune rejection!)
– Do not need to use an embryo
• Cons:
– Cells would still have genetic defects
– One of the pluripotency genes is a cancer gene
– Viruses might insert genes in places we don’t want
them (causing mutations)
The Nobel Prize in Physiology or Medicine 2012

• They opened up a
world of possibilities
What Diseases Do Stem Cells Treat?
Have the Potential to Treat?
Currently Treat Potential to Treat
- Blood Diseases (including immune - Heart Disease
system disorders) - Neurological Diseases (Parkinson’s,
- Genetic metabolic disorders (very Alzheimer’s, Huntington’s & others)
limited/experimental) - Stroke
- Tissue/organ replacement (very - Type 1 Diabetes
limited/experimental) - Macular Degeneration (a common
cause of blindness)
- Cancer
- HIV/AIDS
…and
- Spinal Cord Injury
- Multiple Sclerosis
more!
- ALS (Lou Gehrig’s Disease)
- Liver Disease
 Derivation and Use of Embryonic Stem
Cell Lines
Isolate inner cell mass
Outer cells (destroys embryo)
(forms placenta)
Inner cells
(forms fetus) Culture cells

Day 5-6
Blastocyst “Special sauce”
(largely unknown)

Heart
11/13/2017
repaired
Kidney Heart muscle
Dr. Hariom Yadav
Dr. Hariom Yadav
Dr. Hariom Yadav
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