Evaluation of porcine kidney decellularization protocols and characterization of extracellular matrix
Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesisAlternative title
Utvärdering av decelllulariseringsprotokoll för grisnjure och karaktärisering av extracellulär matrix (Swedish)
The prevalence of chronic kidney disease increases worldwide resulting in more people being affected by the most severe stage; end-stage renal disease. The treatment options available today, hemodialysis and transplantation, can not substitute a fully functioning kidney but the emerging regenerative medicine strategies provide promising treatment alternatives. One strategy is tissues engineering aiming to replace the damaged kidney with a reconstructed one produced by the process of decellularization and recellularization. The success of this strategy is highly dependent on the efficacy of cellular removal and preservation of the kidney extracellular matrix scaffold.
This study evaluated three protocols for decellularization of adult porcine kidney utilizing a combination of different agents; enzymatic agent and ionic, non-ionic and zwitterionic detergents. While the third protocol utilized the ionic detergent sodium dodecyl sulphate, the first and second protocol utilized sodium deoxycholate. The first decellularization protocol utilized a freeze and thaw step additionally. The produced kidney extracellular matrix scaffold was characterized and the findings were that all protocols had a major total protein loss. Although, they all retained the extracellular matrix components elastin, collagen, fibronectin and glycosaminoglycans fairly well.
The results of the study suggests to always use sodium deoxycholate above sodium dodecyl sulphate when choosing an iconic detergent due to being better at retaining the extracellular matrix ultrastructure . In addition, when utilizing the decellularization protocols of this report at least two cycles of decellularization needs to be performed . The main finding of the study was that only the freeze, thaw and sodium deoxycholate protocol for decellularization of adult porcine kidney achieved a satisfactory level of decellularization. This report should be seen as a step in the process of developing a kidney extracellular matrix scaffold optimal for recellularization.
Place, publisher, year, edition, pages
decellularization, kidney transplantation, tissue engineering, extracellulär matrix scaffold, recellularization
Engineering and Technology
IdentifiersURN: urn:nbn:se:kth:diva-173549OAI: oai:DiVA.org:kth-173549DiVA: diva2:853571