Decellularization/Recellularization Flashcards Preview

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Flashcards in Decellularization/Recellularization Deck (6)
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1

What is decell/recell?

process of removing cells from a tissue, followed by reintroducing cells from another individual
can cross species
contingent upon access to the vasculature
revolves around the idea of using the native tissue as a scaffold for new cells

2

What is the technique for recell/decell?

organ is harvested from the donor
tissue is washed with a buffer to remove cells and leave a blank ECM retains much of the cellular structural info
then reseeded with iPSCs or host cells

3

What is the problem for with recell/decell?

there is no standard in the field
uses a combination of physical, enzymatic, and chemical approaches
time scales also have a large degree of variability

4

What are the solutions for recell/decell?

PBS: initial wash of the cells
Trypsin: a protease that breaks down proteins and MHCs
EDTA: chelating agent removes metal ions to shut down native enzymes
NAN3: sodium azide is a antimycotic, toxic to cells, stops fungi growth
CHAPS: mild detergent to break down cell membrane and gentle final wash
Triton X: cell lysis buffer, final step for killing off cells

5

What are common decell assessments?

Gross microscopy w/wo Ponceau Red: does tissue look profused?
Safranin O and Fast Green Stain or microsphere perfusion: chromosomal and nuclear stain/see vascularization
Electron microscopy/fluorescent microsphere perfusion: structural observation
Coupled with DNA quantification

6

What are common recell assessments?

perfused with human cells labeled HUVECs and hMSCs
tests for functionality like tissue mechanics and contractility