NEURAL STEM CELLS TO MODEL NEUROLOGICAL
IPS & NEURAL INDUCTION – Nael Nadif Kasri
Course structure
• Human induced pluripotent stem cells have recently emerges as a novel cellular model to
study the pathophysiology and progress of neurological disorders.
• Aim: Student acquainted with different aspects of hiPS cell research in the context of
neurodevelopmental and neurodegenerative disorders.
• Exam: Proposal about stem cells and brain (70%) and pitch (30%)
o Important: Original/novel, integrating elements of course, study design
o 5 to 7 min pitch
o 5 to 7 min questions
Main learning goals
• Understand and explain current methodologies to generate human induced pluripotent
stem (hiPS) cells
• Distinguish key molecular signalling pathways critical for differentiating hiPS to specific
neuronal cell types
• Understand the advantages and disadvantages of using specific human derived models
• Understand and explain current molecular, cellular and physiological methodologies to
study disease-linked phenotypes
• Apply such methodologies to model neurological disorders in a dish
• Use the newly acquired knowledge to critically evaluate and present a recent manuscript
applying these methodologies to study the molecular and cellular mechanisms underlying a
neurological disorder.
→ Being able to design and write an original project proposal
Stem cells
What are stem cells?
• The foundation cells for every organ and tissue in our bodies
• Stem cells have two key properties:
o The ability to self-renew, dividing in a way that makes copies of themselves
o The ability to differentiate, giving rise to the mature types of cells that make up our organs
and tissues
Stem cell identity and potency
• Totipotent: ability to differentiate into all
possible cell type (i.e. zygote and the first few
cells that result from its division)
• Pluripotent: ability to differentiate into almost
all cell types (i.e. ESCs and cells derived from
the mesoderm, endoderm, and ectoderm germ
layers)
• Multipotent: the ability to differentiate into a
closely related family of cells. (i.e.
hematopoietic (adult) stem cells that can
become red and white blood cells or platelets)
,• Oligopotent: the ability to differentiate into a few cells. (i.e.(adult) lymphoid or myeloid
stem cells)
• Unipotent: the ability to only produce cells of their own type, but have the property of self-
renewal required to be labeled a stem cell. (i.e. (adult) muscle stem cells)
• Embryonic Stem Cells
• Adult Neural Stem Cells
o Neurons don’t divide
o Some new neurons can be formed in HC and LV
• Induced Pluripotent Stem Cells (iPSCs) and Induced
Neuronal Cells (iNS)
Waddington’s epigenetic landscape
The DNA is the same in every cell -> Epigenetic marks
change during differentiation: Activation of genes
Embryogenesis
→ For project ectoderm most important!
One exception: Microglia comes from mesoderm
Also, blood vessels come not from ectoderm
,Embryonic Stem Cells
Mouse Embryonic Stem Cells (mESCs)
Mouse and human ESCs
Promises and limitations of human ESCs
• Advantage: Grown unlimitedly and differentiate to any lineage
• Promises:
o Study human embryogenesis and early development
o Drug screening
o Human disease model
o Personalized regenerative medicine
• Limitations
o Human ESCs from in vitro fertilization
o Ethical issues, destruction of an embryo
→ Induced pluripotent stem cells (iPSCs)
Induced pluripotent stem cells
Nuclear reprogramming: to pluripotency
• Differentiate cells back to stem cells → Reprogramming
-> Molecular prinicple: Epigenetic reprogramming
• Somatic Cell Nuclear Transfer (SCNT)
, • Ectopic expression of transcription factors
o 4 transcription factors (TFs): Oct3/4, Sox2, Klf4 and c-Myc → Yamanaka factors
(OKSM)
-> Add these to somatic cell
Screening of TFs in reprogramming
• Which transcription factors needed for mouse embryonic fibroblast (mMEF) to mouse
induced pluripotent stem cell (miPSC)?...
• Combination of 24 transcription factors (TFs) -> Screening method: put construct in
(promotor which is only active in ES cells) -> Cells on medium they die on -> ES survive
Sequential events of reprogramming
• 3 stages: initiation, maturation and stabilization
• Each stage has its own molecular signature,
which you can use to monitor the stages
• Focus most on IPSC stage
o Once IPSC, different factors are not need
anymore -> Transgene-independent self-
renewal
o Pluripotency
o Once IPSC, cell loses epigenetic memory
o X-reactivation: In every cell one X-
chromosome is deactivated in initiation
stage-> In IPSC stage the deactivated
chromosome become reactivated -> Cells
don’t ‘like’ this…
→ Take this into account by modelling a X-linked disorder
o Telomeres elongation
Human iPSC generation - Characterization
• Cell morphology
• Expression of pluripotency genes
• Differentiation ability: in vitro and in vivo
IPS & NEURAL INDUCTION – Nael Nadif Kasri
Course structure
• Human induced pluripotent stem cells have recently emerges as a novel cellular model to
study the pathophysiology and progress of neurological disorders.
• Aim: Student acquainted with different aspects of hiPS cell research in the context of
neurodevelopmental and neurodegenerative disorders.
• Exam: Proposal about stem cells and brain (70%) and pitch (30%)
o Important: Original/novel, integrating elements of course, study design
o 5 to 7 min pitch
o 5 to 7 min questions
Main learning goals
• Understand and explain current methodologies to generate human induced pluripotent
stem (hiPS) cells
• Distinguish key molecular signalling pathways critical for differentiating hiPS to specific
neuronal cell types
• Understand the advantages and disadvantages of using specific human derived models
• Understand and explain current molecular, cellular and physiological methodologies to
study disease-linked phenotypes
• Apply such methodologies to model neurological disorders in a dish
• Use the newly acquired knowledge to critically evaluate and present a recent manuscript
applying these methodologies to study the molecular and cellular mechanisms underlying a
neurological disorder.
→ Being able to design and write an original project proposal
Stem cells
What are stem cells?
• The foundation cells for every organ and tissue in our bodies
• Stem cells have two key properties:
o The ability to self-renew, dividing in a way that makes copies of themselves
o The ability to differentiate, giving rise to the mature types of cells that make up our organs
and tissues
Stem cell identity and potency
• Totipotent: ability to differentiate into all
possible cell type (i.e. zygote and the first few
cells that result from its division)
• Pluripotent: ability to differentiate into almost
all cell types (i.e. ESCs and cells derived from
the mesoderm, endoderm, and ectoderm germ
layers)
• Multipotent: the ability to differentiate into a
closely related family of cells. (i.e.
hematopoietic (adult) stem cells that can
become red and white blood cells or platelets)
,• Oligopotent: the ability to differentiate into a few cells. (i.e.(adult) lymphoid or myeloid
stem cells)
• Unipotent: the ability to only produce cells of their own type, but have the property of self-
renewal required to be labeled a stem cell. (i.e. (adult) muscle stem cells)
• Embryonic Stem Cells
• Adult Neural Stem Cells
o Neurons don’t divide
o Some new neurons can be formed in HC and LV
• Induced Pluripotent Stem Cells (iPSCs) and Induced
Neuronal Cells (iNS)
Waddington’s epigenetic landscape
The DNA is the same in every cell -> Epigenetic marks
change during differentiation: Activation of genes
Embryogenesis
→ For project ectoderm most important!
One exception: Microglia comes from mesoderm
Also, blood vessels come not from ectoderm
,Embryonic Stem Cells
Mouse Embryonic Stem Cells (mESCs)
Mouse and human ESCs
Promises and limitations of human ESCs
• Advantage: Grown unlimitedly and differentiate to any lineage
• Promises:
o Study human embryogenesis and early development
o Drug screening
o Human disease model
o Personalized regenerative medicine
• Limitations
o Human ESCs from in vitro fertilization
o Ethical issues, destruction of an embryo
→ Induced pluripotent stem cells (iPSCs)
Induced pluripotent stem cells
Nuclear reprogramming: to pluripotency
• Differentiate cells back to stem cells → Reprogramming
-> Molecular prinicple: Epigenetic reprogramming
• Somatic Cell Nuclear Transfer (SCNT)
, • Ectopic expression of transcription factors
o 4 transcription factors (TFs): Oct3/4, Sox2, Klf4 and c-Myc → Yamanaka factors
(OKSM)
-> Add these to somatic cell
Screening of TFs in reprogramming
• Which transcription factors needed for mouse embryonic fibroblast (mMEF) to mouse
induced pluripotent stem cell (miPSC)?...
• Combination of 24 transcription factors (TFs) -> Screening method: put construct in
(promotor which is only active in ES cells) -> Cells on medium they die on -> ES survive
Sequential events of reprogramming
• 3 stages: initiation, maturation and stabilization
• Each stage has its own molecular signature,
which you can use to monitor the stages
• Focus most on IPSC stage
o Once IPSC, different factors are not need
anymore -> Transgene-independent self-
renewal
o Pluripotency
o Once IPSC, cell loses epigenetic memory
o X-reactivation: In every cell one X-
chromosome is deactivated in initiation
stage-> In IPSC stage the deactivated
chromosome become reactivated -> Cells
don’t ‘like’ this…
→ Take this into account by modelling a X-linked disorder
o Telomeres elongation
Human iPSC generation - Characterization
• Cell morphology
• Expression of pluripotency genes
• Differentiation ability: in vitro and in vivo
