Decellularization/Recellularization Flashcards
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
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
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
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
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
What are common recell assessments?
perfused with human cells labeled HUVECs and hMSCs
tests for functionality like tissue mechanics and contractility
