Induced Pluripotent Stem Cells (iPSCs)

and their application in medicine

Dr. Baoqiang Guo

B.guo@ mmu.ac.uk
The outcome of this lecture
By the end of the session you will be able to:
• Describe what is stem cell

• Describe what is pluripotency

• Describe what is pluripotent stem cells,

Including ESCs and iPSCs
• Discuss how to generate iPSCs and their clinical
application potential in medicine
Life starts from a fertilized egg
What is a stem cell?

Character of stem cell:

• Self-renew
• Differentiation
Why are stem cells so important?
Stem cells are different from other mature, specialized cells
of the body , because stem cells can both:
1) Self-renew: Make copies of themselves and
2) Differentiate: Make other types of cells-specialized cells

• ‘Specialized’ or ‘differentiated ‘cells play particular roles

in the body, e.g blood cells, neuron cells, muscles
• Specialized cells can’t divide to make copies of
themselves. This makes stem cells very important.
• The body needs stem cells to replace specialized cells
that are dead or damaged.
Stem cell Jargon

Such as mesenchymal stem cells (MSCs);

Hematopoietic stem cells (HSCs)
 Summary of pluripotency
Pluripotency describes the ability of a cell:

• To develop into the three primary germ cell layers of

the early embryo and therefore into all cells of the
adult body( Endoderm; Mesoderm; Ectoderm).
• Embryonic stem cells (ESCs) and induced pluripotent
stem cells(iPSCs) are characterised by their
pluripotency.
Embryonic stem cell (ESCs) differentiate
into unipotent stem cells (adult stem cells)
ESCs finally differentiate into
unipotent stem cells (adult stem cells)
Embryonic stem cells (ESCs): Where do
we find them?
There is an ethic issue for ESCs research and
application
Discovery of induced pluripotent
stem cells (iPSCs)
Why are iPSCs a ground breaking
discovery –reprogramming?
• Cellular differentiation has long been considered a
permanent one-way process
• Specialized cell types were described as ‘terminally
differentiated’
• Shinya Yamanaka et al. (2006) – demonstrated that
differentiated skin fibroblast cells could be turned
back to a pluripotent embryonic-like state – iPSCs

• iPSCs are pluripotent; unlike ESC, iPSCs are not derived

from the embryo, but instead created from mature cells
in the lab through a process – cellular reprogramming.
The Nobel Prize in Physiology or Medicine 2012

Shinya Yamanaka
and John Gurdon

• Mature cells can be reprogrammed to become pluripotent

iPSCs
• These ground breaking discoveries have completely changed our
view of the development and cellular specialisation
• Equivalent of antibiotics and vaccine in the past century
The 2012 Nobel Prize in Physiology or
Medicine is Announced. (October 9, 2012)
The Nobel Prize in Physiology or Medicine for 2012 was
awarded jointly to Sir John B. Gurdon and Shinya Yamanaka "for
the discovery that mature cells can be reprogrammed to become
pluripotent".

The Nobel Prize recognizes two scientists who

discovered that mature, specialised cells can be
reprogrammed to become immature cells capable of
developing into all tissues of the body. Their
findings have revolutionised our understanding of
how cells and organisms develop.
Comments from Sir Mark Walport, the
director of the Wellcome Trust, said

: "John Gurdon's life has been spent in biology, from

collecting insects as a child to over 50 years at the
laboratory bench. He and Shinya Yamanaka have
demonstrated conclusively that it is possible to turn
back the clock on adult cells, to create all the
specialised cell types in the body.
"Their work has created the field of regenerative
medicine, which has the potential to transform the
lives of patients with conditions such as Parkinson's,
stroke and diabetes.
"This is a wonderfully well-deserved Nobel Prize."
Three milestones lead to the discovery of iPSCs

• John Gurdon: Cloning of frog

• Ian Wilmut and Keith Campbell:

Cloning of Dolly
(reprogramming)

• James Thomson:
Embryonic stem cell lines
derived from human
blastocysts
Cloning of frog by professor John
Gurdon in 1962 ,

He is the first
person to confirm
cell reprogramming
The story of Dolly: Reprogramming

Sir Ian Wilmut - Dolly and Polly ...

From a Orthopaedic doctor to a stem
cell biologist, Nobel prize laureate
24 factors are involved in stem cell
pluripotency
 Yamanaka hypothesis
• Testing of 24 possible pluripotency genes
• Yamanaka factors: Oct4, Sox2, Klf4, c-Myc 4. Cellular
reprogramming
Kazutoshi Takahashi1
Shinya Yamanaka1,2,
The common methods to make iPSCs
cells currently
How are the Yamanaka Factors
introduced into cells?
How do we know that
differentiated cells have been
reprogrammed into iPSCs?
 Testing for Pluripotency

1. Morphology (cell shape)

2. Genomics (types of genes expressed)

3. Function (ability to differentiate)

Morphology
• Change in morphology of the cells*
• Example: skin cells grow as flattened cells in a
dish; however, as they become reprogrammed, the
iPSC grow in round clumps known as colonies
Negative SSEA4

SSEA4
SSEA4
Tra-1-60

Tra-1-60 SSEA4-Tra-1-60

SSEA4
SSEA4

Tra-1-60 Tra-1-60
Expression of pluripotency
markers
• Pluripotency markers
• Genes that are only expressed in
pluripotent stem cells and not in other cell
types.
• Its expression resembles a molecular
signature that lets scientists know that
cells have been reprogrammed to a
pluripotent state.
Pluripotency markers
Genes upregulated in ES
and/or iPS cells.
Genes in group I are
genes upregulated in ES
cells and iPS cells.
Genes in group II are
upregulated more in ES
cells, iPS-MEF4-7,
and iPS-MEF10-6 than in
iPS-MEF3 cells.
Genes in group III are
upregulated more in
ES cells than in iPS cells.
Lists of genes are
shown in Tables S3–S5.
Function:
Ability to differentiate into cell types of the 3
embryonic germ layers both in vitro and in vivo
 What are the potential applications of
iPSCs?
• A good research tool for biology study.
• iPSCs technology is powerful method for human
diseases modelling
• IPSCs are useful tools for drug development .
• iPSCs derived cardiomyocytes, lever cells and
kidney cells for toxicology
• Scientists hope to use them in transplantation
medicine.
Summarise the application of iPScs
 iPSCs therapy for heart failure
1. Generation of safe iPSCs 2. Effective cardiac differentiation

5. Careful 4. Effective and safe 3. Large scale culture and

observation of transplantation of purification of cardiomyocyte
arrhythmogenicity purified cardiomyocyte
Kyoto Univ. performs world's 1st iPS cell transplant for Parkinson's
NOVEMBER 09, 2018 18:45 JST
KYOTO (Kyodo) -- Kyoto University said Friday it has conducted the world's
first transplant of induced pluripotent stem cells to treat Parkinson's disease.
Nerve cells created from the artificially derived stem cells were transplanted
into the brain of a patient in his 50s in October in a treatment which
researchers hope to develop into a method covered by Japan's health
insurance.
Insulin-producing pancreatic beta cells (green)

Caption: Insulin-producing pancreatic beta cells (green) derived from human

embryonic stem cells that have formed islet-like clusters in a mouse. The red
cells are producing another metabolic hormone, glucagon, that regulates blood
glucose levels. Blue indicates cell nuclei.
Credit: Photo by B. D. Colen/Harvard Staff; Image courtesy of Doug Melton
Revolutionizing Regenerative Medicine
with Mass Manufacturing of Induced
Pluripotent Stem Cells (iPSCs)

Closed Automatic iPSC Mass Manufacturing System

The outcome of this lecture

By the end of the session you will be able to:

• Describe what is stem cell

• Describe what is pluripotency

• Describe what is pluripotent stem cell,

Including ESCs and iPSCs
• Discuss how to generate iPSCs and their clinical
application potential in medicine
 Reading list

1.Takahashi, K. & Yamanaka, S. Cell 126, 663–676 (2006).

2. Takahashi, K. et al. Cell 131, 861–872 (2007)
2. Penelope J. et al. Cell Stem Cell 16, 269–274( 2015)
Thank you for
attending the lesson