Read the summary and the most important questions on Introduction to Stemcells

• 1 Day 1: Embryonic stem cells, pluripotency and reprogramming

• 1.1 From embryo to embryonic stem cells (Roelen)

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• Oocyte (egg cell)

  Large immotile cell.

• Which tissues can a pluripotent stem cells differentiate towards?

  Endoderm, mesoderm, germ cells (NO yolk sack and placenta).

• 1.3 Reprogramming, induced pluripotent stem cells and application (Jaenisch)

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• Sporadic, late onset diseases

  • Common and therefore medically the most relevant
  • Lack a clear genetic basis
  • Generally display a more subtle phenotype   

• Therapeutic applications of iPSCs?

  • Can be used to generate desired cell types for individual patients
  • Can be used to model disease, facilitating drug discovery.
  • Patient-derived iPSCs could potentially be corrected ex vivo to be transplanted back into the patient to replenish lost/damaged cells.   

• Major issue with iPSC-derived cells

  They often remain poorly matured

• How can we manipulate the human (epi)genome to rectify disease-causing variants?

  • Gene targeting
    
  • CRISPR/Cas9-bases gene editing

• CRISPR/Cas9-based gene editing

  • Efficient method to generate/correct mutations, based on HR on nonhomologous end joining (NHEJ)
  • Involves a guide RNA (for homology search) and Cas9 (cleaves DNA).
  • Can introduce multiple genetic modifications at a time.
  • Efficient and rapid technique.
  • Does not require extensive selection process.    

• Two main types of gene therapies?

  • Somatic therapy
    
  • Germline therapy

• 2 Day 2: Epigenetics and flavors of pluripotency

• 2.1 Principles of epigenetics in stem cells (Lopes)

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• Association of ATP-dependent complexes

  • E.g. polycomb and trithorax proteins
  • Change the distance between nucleosomes, which results in altered accessibility 

• Association with non-coding RNAs

  • Binding of non-coding RNAs to the DNA blocks transcription
  • E.g. X-inactive specific transcript (XIST) and TSIX
  •  XIST and TSIX can neutralize each other