Paper: Regeneration of the entire human epidermis using transgenic stem cells

9 important questions on Paper: Regeneration of the entire human epidermis using transgenic stem cells

What are the four degrees of burn wounds?

  1. Sunburn
  2. Blistering
  3. Epidermal defects, going into the dermis --> hair follicles are lost
  4. complete loss of skin

What is the golden standard in therapy for therapy of burn wounds?

Autologous split skin mesh grafts: shave of a piece of patients skin, and put holes in it, so you can stretch it

What are the pros and cons of autologous split skin mesh grafts?

Pros: no rejection because own skin, quite fast (this is important because having no skin for a long time can lead to dehydration and injections (1st line of defesce) and further problems, which are life-threatening).

Cons: you create even more wounds, sometimes there is not enough healthy skin (for example when the whole body is burned).
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Explain the principle of cultured epithelial autografts

A patient skin biopsy is taken, and epidermal cells are isolated. These are plated on 3T3 feeder cells, and keratinocytes are amplified by subculturing. An epithelial sheet is formed, and enzymatic detachment dispase of thermolysine is added. The cells are then transferred and attached to a backing material, and transplantation can be performed.

What are the two types of heritable skin diseases?

Cornification disorders and blistering skin disease.

What are the three types of epidermolysis bullosa?

  1. EB simplex: autosomal dominant (mutation in keratin 5/14)
  2. Junctional EB: autosomal dominant (mutations in Laminin, Collagen 17)
  3. Dystrophic EB: autosomal dominant or recessive (mutations in collagen 7)

To what manifestations can the three different types of EB lead?

EB simplex: superficial blisters between epidermis
Junctional EB: separation of the dermis & epidermis
Dystrophic EB: very deep dermal cleavage

What are the two treatment options for junctional EB?

  1. Gene addition --> by viral transduction
  2. Gene correction --> by Crispr-Cas9

Explain the principle of correction of junctional EB by using iPSCs

  • Take mutant somatic cells from patient with heritable disease
  • Make iPSCs
  • Correct the gene defect in vitro (large cell population)
  • generate keratinocytes or fibroblasts (full differentiation)
  • transplant an epidermal sheet of corrected cells to the patient

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