期刊
AMERICAN JOURNAL OF TRANSPLANTATION
卷 15, 期 1, 页码 64-75出版社
WILEY
DOI: 10.1111/ajt.12999
关键词
animal models: murine; bioengineering; kidney biology; stem cells; basic (laboratory) research; science; regenerative medicine; tissue; organ engineering
资金
- Zell Family Foundation
- Excellence in Academic Medicine Act through Illinois Department of Healthcare and Family Services
- Northwestern Memorial Foundation Dixon Translational Research Grants Initiative
- American Society of Transplant Surgeon's Faculty Development Grant
- NIDDK [R01 DK050141]
- NIGMS [K12 GM088021]
- Dialysis Clinic, Inc.
- Northwestern University
- Northwestern University Mouse Histology and Phenotyping Laboratory
- Cancer Center Support Grant (NCI) [CA060553]
- NCI CCSG [P30 CA060553]
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 retentionindicating 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 3h 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. The authors validate an optimal detergent-based protocol for decellularization of rodent whole-kidney scaffolds, showing that decellularized scaffolds retain an intact vasculature that can be transplanted or re-endothelialized, wand that the scaffold supports proliferation and tubule formation by human renal cortical tubular epithelial cells.
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