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(keynote): Induced pluripotent stem cells (ipSC)

23 important questions on (keynote): Induced pluripotent stem cells (ipSC)

What is the developmental significance of a fertilized egg being initially referred to as totipotent and later as pluripotent?

In the early stages after fertilization, the migrating fertilized egg is termed totipotent, signifying its ability to differentiate into any cell type, including embryonic and extraembryonic cells. As development progresses, the cells become pluripotent, maintaining the capacity to differentiate into various cell types but losing the ability to form all cell types, as observed in the totipotent stage.

Give some examples of stem cells and where in the body we can find them.

  • Hematopoeitic stem cells: bone marrow
  • Mesenchumal stem cells: bone marrow, adipose tissue, connective tissue
  • Neural stem cells: Nervous system
  • Embryonic stem cells: inner cell mass of a developing embryo
  • Induced pluripotent stem cells, generated by reprogramminga dult cells such as skin or blood

What is pluripotent differentiation, and from which embryonic structure are embryonic stem cells derived during the blastocyst stage?









Pluripotent differentiation refers to the ability of stem cells to differentiate into various cell types derived from all three germ layers (ectoderm, endoderm, and mesoderm). Embryonic stem cells are derived from the inner cell mass of the blastocyst stage embryo, a hollow structure formed during early embryonic development.
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Explain: totipotent: pluripotent: multipotent



Totipotent: Can differentiate into any cell type, including extraembryonic tissues (e.g., zygote).
Pluripotent: Can differentiate into cells of all three germ layers (ectoderm, mesoderm, endoderm), as seen in embryonic stem cells.
Multipotent: Can differentiate into a limited range of cell types within a specific lineage or tissue, more specialized than pluripotent cells.

Where are the two main regions in the brain where adult neural stem cells are found, and what is the role of these regions in the ongoing neurogenesis of the adult brain?

The two main regions in the brain where adult neural stem cells are found are the subventricular zone, which lines the lateral ventricles, and the hippocampus, a structure associated with learning and memory. These regions house pools of adult neural stem cells that contribute to the ongoing neurogenesis, or formation of new neurons, in the adult brain.

Tell something about the potency and regenerative potential of ESCs and adult/tissue stem cells.

  1. Embryonic Stem Cells (ESCs):
    • Potency: Pluripotent, meaning they can differentiate into cells of all three germ layers (ectoderm, mesoderm, endoderm).
    • Promise for Regenerative Medicine: ESCs show high promise for regenerative medicine due to their remarkable differentiation potential, making them suitable for a wide range of tissue and organ repair.
  2. Adult/Tissue Stem Cells:
    • Potency: Generally multipotent, meaning they can differentiate into a limited range of cell types within a specific tissue or lineage.
    • Regenerative Potential: While not as versatile as ESCs, adult/tissue stem cells play a crucial role in tissue maintenance and repair in their respective organs or tissues.

Pluripotent stem cells can be generated through a process known as nuclear reprogramming. This involves altering the gene expression profile of a differentiated cell, typically a somatic cell, to reset it to a pluripotent state. The most notable method of nuclear reprogramming is the induction of pluripotency, leading to the creation of induced pluripotent stem cells (iPSCs).
Name 3 methods of Inducing Pluripotency in Somatic Cells:

  • Nuclear transfer
  • Cell fusion
  • Defined factors

Explain how defined factors work

Defined factors, consisting of specific transcription factors, are introduced into somatic cells to induce reprogramming. This process triggers changes in the gene expression profile of the somatic cell, ultimately resetting it to a pluripotent state. The introduction of these defined factors is associated with the creation of induced pluripotent stem cells (iPSCs). This method allows for the generation of pluripotent stem cells without the ethical concerns associated with embryonic stem cells.

How to test iPSCs for pluripotency?

  • It needs to give rise to more differentiated cell types.
  • If these pluripotent stem cells are able to have a mesoderm, ectoterm and endoderm layer.

Name 3 ways to identify and characterize induced pluripotent stem cells.

Embryonic Stem Cell-Like Properties:

  • iPSCs share similarities with embryonic stem cells (ESCs).
  • They have the potential to differentiate into cell types from all three germ layers.
Pluripotency Markers:
  • iPSCs express specific markers indicative of pluripotency.
  • Alkaline phosphatase is one such marker, but it is not exclusive to pluripotent cells.
Karyotype and Chromosome Integrity:
  • Assessment of the karyotype ensures the normal number and structure of chromosomes.
  • This step is crucial to exclude cells with DNA damage or abnormalities.









How did Gurdon and Yamanaka's groundbreaking discoveries reshape our understanding of development and cellular specialization, challenging the notion that mature cells are permanently confined to their specialized states?

Gurdon's demonstration that the DNA of mature cells in frogs contains all the information for developing all cell types, and Yamanaka's discovery of reprogramming intact mature cells into stem cells in mice, have collectively transformed our perspective on development and cellular specialization. These findings challenged the belief that mature cells are permanently confined to their specialized states, highlighting the potential for reprogramming and the plasticity of cellular identity.

The principle of cellular reprogramming via somatic cell nuclear transfer:

Somatic cell nuclear transfer involves transferring the nucleus of a somatic cell into an enucleated egg cell, triggering reprogramming and generating pluripotent stem cells.

The principle of cellular reprogramming via induced pluripotent stem cell technology

Induced pluripotent stem cell (iPSC) technology involves introducing specific transcription factors into somatic cells, resetting their gene expression profile to induce pluripotency.

How do you know a cell is pluripotent?

Pluripotent cells express specific markers indicative of pluripotency. However, additional assessments, such as examining their ability to differentiate into cells of all three germ layers and ensuring chromosome integrity (karyotype analysis), are crucial to confirm their pluripotent nature.

What are examples of monogenic disorders and complex genetic disorders?

Monogenic disorders: metabolic disorders, leukodystrophies, epilepsy.
Complex genetic disorders: Schizophrenia and Depression

What are the limitations associated with using postmortem tissue and animal models in research?

Current research based on postmortem tissue is limited by the fact that it primarily reflects the end-stage of diseases. Additionally, using animal models presents a disadvantage, particularly in the study of neuropsychiatric disorders, as these conditions are often human-specific, and findings may not fully translate to human pathology.

Schizophrenia patients
  • Prevalence ...
  • Onset usually dusring .....
  • .... societal burden (cost, mortality, emotional)
  • Heriability

iPSC generation of schizophrenia patients
  • Prevalence 1%
  • Onset usually dusring adolescence
  • High societal burden (cost, mortality, emotional)
  • Heriability 81%

What signals contribute to the formation of stem-cell precursors in the embryonic neural tube?

Signals from the ventral floor plate (e.g., Sonic Hedgehog or SHH) and roof plate (Bone Morphogenetic Proteins or BMPs) contribute to the formation of stem-cell precursors in the embryonic neural tube.

What markers are used to identify the presence of both glutamatergic and GABAergic synapses in a culture of cortical neurons?

Markers such as VGAT (GABAergic presynaptic marker) and VGLUt1 (glutamatergic presynaptic marker) are used to identify the presence of both glutamatergic and GABAergic synapses in a culture of cortical neurons.

How do neurons derived from Tuberous Sclerosis Complex (TSC) patients differ in terms of activity?

Neurons derived from Tuberous Sclerosis Complex (TSC) patients show increased activity, contributing to the major problem of epilepsy associated with this condition.

Describe the process of human astrocyte generation using induced pluripotent stem cells (iPSCs).

Human astrocyte generation involves the use of common glia precursors, embryoid bodies (EBs), neural epithelial spheres (NES), early glial precursor cells (GPCs), and late GPCs to produce astrocytes, which play a supportive role in neuronal function.

Name three applications of iPSC technology, emphasizing its versatility in different fields.

Three applications of iPSC technology include drug development (patient-specific cells for drug screening), cell replacement therapy (corrected patient-specific iPSCs for transplantation), and disease modeling (in vitro modeling of complex genetic disorders).

How can iPSC technology contribute to neuroscience research, and provide an example of its application in pre-clinical research?

iPSC technology contributes to neuroscience research by providing personalized models for drug testing, cell replacement therapies, and disease modeling. An example in pre-clinical research involves using transcription factors to identify cortical GABAergic interneurons in germinal layers like ganglionic eminences.

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Developing brain

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