Induced Pluripotent Stem Cells
Generation and their Therapeutic
M.Sc IInd YEAR BMB
DEPT. OF BIOCHEMISTRY & MOLECULAR BIOLOGY
DEPT. OF BMB
• Induced pluripotent stem cells (iPS cells) are
somatic cells that have been reprogrammed to
a pluripotent state by the introduction of
• iPS cells are similar to embryonic stem (ES)
markers and the capacity to develop
• Yamanaka and Sir John Gurdon (2012) were
awarded the Nobel Prize in Physiology or
Medicine "for the discovery that mature cells
can be reprogrammed to become pluripotent."
DISCOVERY OF iPS CELLS
• The human ES cells derived from human blastocysts
were first established by Thomson et al.
• The iPS cells were first established in 2006 by Takahashi
and Yamanaka by the retrovirus-mediated transduction
of four transcription factors (c-Myc, Oct3/4, SOX2, &
Klf4) into mouse fibroblasts.
• Human iPS cells were established in 2007, by the
transduction of either the same set of transcription
factors or another set of transcription factors (Oct3/4,
SOX2, Nanog, Lin28) into human fibroblast.
• Human iPS cells have been reported to be established
from skin fibroblasts, keratinocytes, and mobilized
CD34+ hematopoietic stem/progenitor cells
differentiated T cells from peripheral blood.
• Expression is essential for the development of
the inner cell mass (ICM) in vivo, the derivation
of ES cells and the maintenance of a pluripotent
• Transcription factor involved in the self-renewal
of ES cells. It has an important role in
heterodimerizes in a complex with Oct4.
• Pleiotropic transcription factor that has been
linked to several cellular functions, including cell
differentiation and metabolism.
• Expressed in a variety of tissues, including the
epithelium of the intestine, kidney and the skin.
• Forced overexpression of Klf4 in ES cells inhibits
differentiation in erythroid progenitors, suggesting a role
for this factor in ES-cell function.
• Transcription factor that was involved in maintaining EScell self-renewal and pluripotency.
• Nanog-deficient ES cells completely lose their selfrenewal capability, differentiating into extra-embryonic
• Expressed in ES cells and during early embryogenesis
but its expression becomes restricted to several tissues
during late embryogenesis and adult life.
iPSC has been generated from
Mouse (Yamanaka et al., 2006)
Humans (Yamanaka et al., 2007)
Rhesus monkey (Liu et al., 2008)
Rats (Liao et al., 2009; Li et al., 2009)
Canine (Shimada, H. et al, 2010)
Porcine ( Esteban, M. A. et al., 2009)
Marmoset (Wu, Y. et al., 2010)
Rabbit (Honda, A. et al., 2010)
Equine (Kristina Nagy et al., 2011 )
Avian (Lu et al., 2011)
iPSC Therapeutic Applications (Cont)
Monogenic and Polygenic Diseases
• Generation of iPS cells from patients with a variety
of genetic diseases with either Mendelian or
complex inheritance has been described.
ADA-SCID, Gaucher disease type
III, Huntington disease, juvenile-onset type 1
diabetes mellitus , Down syndrome.
• Patient-specific fibroblasts offer a unique
opportunity for studying and modeling the effects
of specific gene defects on human neuronal
development in vitro and other potential therapies
for the relevant neurogenetic disorders.
• Patient-derived iPS cells could be subsequently
differentiated in vitro into dopaminergic neurons
for treatment of Parkinson's disease.
iPSC Therapeutic Applications (Cont)
Degenerative Cardiac Diseases
• iPS cell based cell replacement therapy is currently
generating a great deal of interest in the treatment of
ischemic heart diseases since iPS cells are capable of
Obstacles in therapeutic application of iPSCs in humans
(i) Use of harmful oncogenes as part of the reprogramming
(ii) Use of viral vectors for gene delivery that carry the risk of
(iii) Low efficiency and slow kinetics of reprogramming.
(iv) Lack of robust and reliable differentiation protocols for
human iPS cells.
• With an aging population, ever-increasing costs of
healthcare and a decrease in new drug
discoveries, new and innovative ways of accelerating
and streamlining human disease study and treatment
• Human iPSC technology offers an unprecedented
opportunity to develop patient-specific and diseasespecific iPSC lines to be used for regenerative
therapies, disease study, drug safety and discovery.
• The potential of iPS cell technology is tremendous, but
this technology is still in its infancy.
• To realize the full application of iPS cells, it will be
essential to improve the methodologies for iPS cell
generation and to precisely evaluate each clone and
sub clone of iPS cells for their safety and efficacy .
• Takahashi K, Yamanaka S. Induction of pluripotent stem
cells from mouse embryonic and adult fibroblast cultures by
defined factors. Cell. 2006;126:663–676.
• Shinya Yamanaka & Helen M. Blau. Nuclear reprogramming
to a pluripotent state by three approaches iPS cells: A
source of cardiac regeneration Yoshinori Yoshida, Shinya
• Giovanni Amabile and Alexander Meissner .Induced
pluripotent stem cells: current progress and potential for
Sun,Reginald Liew. Clinical applications of patient-specific
induced pluripotent stem cells in cardiovascular medicine
• Yoshida Y, Yamanaka S. iPS cells: a source of cardiac
regeneration. J Mol Cell Cardiol 2011;50:327e32
• Alejandra M. Vitalea, Ernst Wolvetangb, Alan MackaySima.Induced pluripotent stem cells: A new technology to study