The impact of decellularization agents on renal tissue extracellular matrix.
The combination of patient-specific cells with scaffolds obtained from natural sources may result in improved regeneration of human tissues. Decellularization of the native tissue is the first step in this technology. Effective decellularization uses agents that lyse cells and remove all cellular materials, leaving intact collagenous extracellular matrices (ECMs). Removing cellular remnants prevents an immune response while preserving the underlying structure. In this study, the impact of five decellularization agents (0.1 N NaOH, 1% peracetic acid, 3% Triton X-100, 1% sodium dodecyl sulfate (SDS), and 0.05% trypsin/EDTA) on renal tissue was examined using slices of porcine kidneys. The NaOH solution induced the most efficient cell removal, and resulted in the highest amount of cell viability and proliferation after recellularization, although it also produced the most significant damage to collagenous fiber networks, glycosaminoglycans (GAGs) and fibroblast growth factor (FGF). The SDS solution led to less severe damage to the ECM structure but it resulted in lower metabolic activity and less proliferation. Peracetic acid and Triton X-100 resulted in minimum disruption of ECMs and the most preserved GAGs and FGF. However, these last two agents were not as efficient in removing cellular materials as NaOH and SDS, especially peracetic acid, which left more than 80% of cellular material within the ECM. As a proof of principle, after completing the comparison studies using slices of renal ECM, the NaOH process was used to decellularize a whole kidney, with good results. The overall results demonstrate the significant effect of cell lysing agents and the importance of developing an optimized protocol to avoid extensive damage to the ECM while retaining the ability to support cell growth.
Department of Chemical Engineering, Brigham Young University, Provo, UT, USA.
Nafiseh
N
Poornejad
Department of Chemical Engineering, Brigham Young University, Provo, UT, USA.
Lara B
LB
Schaumann
Department of Genetics and Biotechnology, Brigham Young University, Provo, UT, USA.
Evan M
EM
Buckmiller
Department of Chemical Engineering, Brigham Young University, Provo, UT, USA.
Nima
N
Momtahan
Department of Physiology and Developmental Biology, Brigham Young University, Provo, UT, USA.
Jason R
JR
Gassman
Department of Physiology and Developmental Biology, Brigham Young University, Provo, UT, USA.
Ho Hin
HH
Ma
Department of Biology, Brigham Young University, Provo, UT, USA.
Beverly L
BL
Roeder
Department of Physiology and Developmental Biology, Brigham Young University, Provo, UT, USA.
Paul R
PR
Reynolds
Department of Chemical Engineering, Brigham Young University, Provo, UT, USA cook@byu.edu.
Alonzo D
AD
Cook
D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D002474:Cell-Free System / Q000737:chemistry*; D002478:Cells, Cultured; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005109:Extracellular Matrix / Q000737:chemistry* / Q000648:ultrastructure*; D007668:Kidney / Q000737:chemistry* / Q000648:ultrastructure; D008422:Materials Testing; D013501:Surface-Active Agents / Q000737:chemistry*; D013552:Swine; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods; D054457:Tissue Scaffolds
D013501:Surface-Active Agents
Decellularization; alkaline/acidic; ionic/non-ionic detergent; renal tissue