Adobe Systems
Induced pluripotent stem cells (iPSC)
1
Induced pluripotent stem cells (iPSC)
Kseniya Bobryshava and Marina Vasenko

Adobe Systems
Induced pluripotent stem cells (iPSC)
2
Pluripotent stem cells
̶Embryonic stem cells
̶We can find them in blastocyst (5-9 days)
̶Are able to differentiate to 220 types of cells
̶Have the ability to reproduce indefinitely in culture, and incorporate into host embryos

Adobe Systems
Induced pluripotent stem cells (iPSC)
3
Multipotent stem cells
̶Adult or somatic stem cells that can differentiate into numerous related cells
̶Differentiate from endoderm
̶Classical example: hematopoietic stem cells
̶Most blood stem cells can be found in the bone, but the small number can be found in the
bloodstream
̶

Adobe Systems
Induced pluripotent stem cells (iPSC)
4
Unipotent stem cells
̶These stem cells can produce only one cell type.
̶Have the property of self-renewal that distinguishes them from non-stem cells.
̶Examples of a unipotent stem cell are a germ line stem cell (producing sperm) and an epidermal
stem cell (producing skin).

Adobe Systems
Induced pluripotent stem cells (iPSC)
5
Embryonic stem cells vs adult stem cells
You call it
life?

Adobe Systems
Induced pluripotent stem cells (iPSC)
6
Shinya Yamanaka
̶Shinya Yamanaka was born on September 4, 1962.
̶As a young man Yamanaka was a resident in orthopedic surgery.
̶ Yamanaka states in his interviews that he decided to devote himself to research after his
father’s death. He was devastated about the fact, that he was a doctor, but he couldn’t help his
own father.
̶ The next 10 years Yamanaka spent in his PhD studies and postdoc work and in 2006 he presented
induced pluripotent stem cells for the first time.

Adobe Systems
Induced pluripotent stem cells (iPSC)
7
Yamanaka’s experiment
̶24 transcription factors
̶The four Yamanaka factors are Oct4 (O), Sox2 (S), Klf4 (K), and c-Myc (M).
̶Yamanaka and his team used retroviral vectors to deliver the reprogramming factors into the cells.
This process is often referred to as transduction.
̶

Adobe Systems
Induced pluripotent stem cells (iPSC)
8
Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined
Factors
̶After successful transduction cells became resistant to G418 (Geneticin), antibiotic that blocks
polypeptide synthesis by inhibiting the elongation step in both prokaryotic and eukaryotic cells.
̶The reprogrammed cell was determined as an iPSC if it had round shape, large nucleoli, and scant
cytoplasm and also expressed embryonic stem cell markers.
̶To determine the optimal combination of transcription factors, the researchers took one factor at
a time and monitored the difference in the number and morphology of outstanding colonies.
̶

Adobe Systems
Induced pluripotent stem cells (iPSC)
9
Disadvantages of iPSC
̶The initial idea of using IPSC for creating organoid, tissue or whole organs was really ambitious
but in reality had a lot of issues.
̶The process is time-consuming and is manually done by well-trained technicians.
̶Since each technician can only comfortably prepare cells for a few patients in one experiment
cycle, large numbers of technicians will be required for large-scale production. This raises the
production cost and limits the production capacity.

Adobe Systems
Induced pluripotent stem cells (iPSC)
10
Disadvantages of iPSC
̶Additionally, manual operation brings in human errors, which can result in production failure.
̶Only one patient's cells can be handled in each clean room at any given time to avoid
cross-contamination leads to limited production capacity per cGMP facility.
̶The risk of viral DNA mess with genome
̶The risk of teratoma formation or appearance of cancer (one of Yamanaka’s factors is onco factor)
Reducing Human Errors in the Lab

Adobe Systems
Induced pluripotent stem cells (iPSC)
11
Heart Diseases
̶Many heart diseases manifest due to changes in the mechanical strain of cardiac tissue.
̶Direct transplantation of cell sheets derived from iPSCs cardiomyocytes onto the damaged heart
areas has been proven to improve cardiac function in damaged hearts. Unfortunately, this method has
still some issues like low  percentage of injected cells successfully engrafting in the host heart
or occurrence of ventricular arrhythmia.
̶Nowadays iPSCs are mostly used for studying different diseases in vivo like Long QT syndrome,
hypertrophic/dilated cardiomyopathy or arrhythmogenic cardiomyopathy.
Obsah obrázku text, snímek obrazovky, diagram Popis byl vytvořen automaticky Obsah obrázku snímek
obrazovky, kosti Popis byl vytvořen automaticky

Adobe Systems
Induced pluripotent stem cells (iPSC)
12
Neurological Disorders
Obsah obrázku text, snímek obrazovky, řada/pruh, číslo Popis byl vytvořen automaticky
̶There are many neurological diseases, that potentially could be cured thanks to iPSCs.
̶ Parkinson's disease is associated with gene mutations such as SNCA, LRRK2, VPS35, Parkin, PINK1,
DJ-1, and CHCHD2. Researchers successfully generated iPSC lines from patients with all different
mutations
  allowing for the study of disease
  mechanisms, drug testing, and
  potential personalized therapies.
̶

Adobe Systems
Induced pluripotent stem cells (iPSC)
13
Neurological Disorders
̶In case of Alzheimer's disease iPSCs were used not only for studying and treating this disorder,
but also for searching a preventing method for amyloid-beta aggregation and plaque formation of
neurons to improve neuronal viability.
̶
Obsah obrázku text, snímek obrazovky Popis byl vytvořen automaticky

Adobe Systems
Induced pluripotent stem cells (iPSC)
14
Ophthalmic Diseases
̶Stem cell treatments appear to offer hope to people with forms of age-related macular degeneration
(AMD), retinitis pigmentosa (RP), and Stargardt disease.
̶In 2013, an ophthalmologist Masayo Takahashi and her team made iPS cells from the skin cells of
two people with age-related macular degeneration and used them to create sheets of retinal pigment
epithelium (RPE) cells for a clinical trial.
̶In 2014, they implanted these sheets into the right eye of a woman in her seventies, effectively
halting macular degeneration and improving her vision.
Human-induced pluripotent stem cells-derived retinal pigmented epithelium, a new horizon for
cells-based therapies for age-related macular degeneration | Stem Cell Research & Therapy | Full
Text

Adobe Systems
Induced pluripotent stem cells (iPSC)
15
Diabetes
̶Patient tissue biopsies can be reprogrammed into iPSCs and differentiated into stem cell-derived
islets (SC-islets) that contain insulin-secreting β cells. These SC-islets have applications in
disease modeling, gene editing, and cell therapy in animal models. They also provide a source for
autologous cell replacement therapy for patients with diabetes.
̶

Adobe Systems
Induced pluripotent stem cells (iPSC)
16
Personalized medicine - treatment of leukemia
̶Usually the patients have to spend months in the hospital just to find the right medicine for
them.
̶Many of these treatments have a lot of negative side effects that may make the patient’s condition
even worse.
̶In the case of leukemia the mutation appears only in diseased cells so we can isolate patient
blood cells, reprogram them into stem cells, and then observe how the treatment affects them.

Adobe Systems
Induced pluripotent stem cells (iPSC)
17
When modern methods meet together…
̶Many diseases are associated with mutations in the genome of all somatic cells, for instance,
neuropathic pain.
̶
̶CRISPR in iPSC can be used to generate healthy cells.
̶

Adobe Systems
Induced pluripotent stem cells (iPSC)
18
HLA stock project
̶iPS cells are being made to express the most frequent HLA in Japan.
̶Currently, a total of 27 iPS cell lines made from 7 donors who together are homozygous for 4 HLA
types have been prepared. These lines cover approximately 40% of the Japanese population.
̶The different lines give versatility in HLA typing and differentiation capacity for different
diseases.
̶Some of these cell lines have already been used in iPS cell-based clinical cell therapies.

Adobe Systems
Induced pluripotent stem cells (iPSC)
19
HLA stock project

Adobe Systems
Induced pluripotent stem cells (iPSC)
20
Thank you for attention
Obsah obrázku text, kreslené, ilustrace, klipart Popis byl vytvořen automaticky