Optimization and critical evaluation of decellularization strategies to develop renal extracellular matrix scaffolds as biological templates for organ engineering and transplantation.
The ability to generate patient-specific cells through induced pluripotent stem cell (iPSC) technology has encouraged development of three-dimensional extracellular matrix (ECM) scaffolds as bioactive substrates for cell differentiation with the long-range goal of bioengineering organs for transplantation. Perfusion decellularization uses the vasculature to remove resident cells, leaving an intact ECM template wherein new cells grow; however, a rigorous evaluative framework assessing ECM structural and biochemical quality is lacking. To address this, we developed histologic scoring systems to quantify fundamental characteristics of decellularized rodent kidneys: ECM structure (tubules, vessels, glomeruli) and cell removal. We also assessed growth factor retention--indicating matrix biofunctionality. These scoring systems evaluated three strategies developed to decellularize kidneys (1% Triton X-100, 1% Triton X-100/0.1% sodium dodecyl sulfate (SDS) and 0.02% Trypsin-0.05% EGTA/1% Triton X-100). Triton and Triton/SDS preserved renal microarchitecture and retained matrix-bound basic fibroblast growth factor and vascular endothelial growth factor. Trypsin caused structural deterioration and growth factor loss. Triton/SDS-decellularized scaffolds maintained 3 h of leak-free blood flow in a rodent transplantation model and supported repopulation with human iPSC-derived endothelial cells and tubular epithelial cells ex vivo. Taken together, we identify an optimal Triton/SDS-based decellularization strategy that produces a biomatrix that may ultimately serve as a rodent model for kidney bioengineering.
Comprehensive Transplant Center, Feinberg School of Medicine, Northwestern University, Chicago, IL; Department of Surgery, Feinberg School of Medicine, Northwestern University, Chicago, IL; Servei Cirurgia HepatoBilioPancreatica i Trasplantaments, Hospital Universitari Vall Hebron, Universitat Autonoma de Barcelona, Barcelona, Spain.
M
M
Caralt
J S
JS
Uzarski
S
S
Iacob
K P
KP
Obergfell
N
N
Berg
B M
BM
Bijonowski
K M
KM
Kiefer
H H
HH
Ward
A
A
Wandinger-Ness
W M
WM
Miller
Z J
ZJ
Zhang
M M
MM
Abecassis
J A
JA
Wertheim
D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D003902:Detergents / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology*; D005109:Extracellular Matrix / Q000502:physiology*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D007684:Kidney Tubules / Q000098:blood supply / Q000187:drug effects / Q000502:physiology*; D008297:Male; D016377:Organ Transplantation / Q000592:standards*; D010477:Perfusion; D051381:Rats; D017207:Rats, Sprague-Dawley; D023822:Tissue Engineering; D054457:Tissue Scaffolds
D003902:Detergents
animal models: murine, bioengineering, kidney biology, stem cells; basic (laboratory) research/science; regenerative medicine; tissue/organ engineering