Optimization and critical evaluation of decellularization strategies to develop renal extracellular matrix scaffolds as biological templates for organ engineering and transplantation

M. Caralt, J. S. Uzarski, S. Iacob, K. P. Obergfell, N. Berg, B. M. Bijonowski, K. M. Kiefer, H. H. Ward, A. Wandinger-Ness, W. M. Miller, Z. J. Zhang, M. M. Abecassis, J. A. Wertheim*

*Corresponding author for this work

Research output: Contribution to journalArticle

105 Scopus citations

Abstract

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 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.

Original languageEnglish (US)
Pages (from-to)64-75
Number of pages12
JournalAmerican Journal of Transplantation
Volume15
Issue number1
DOIs
StatePublished - Jan 1 2015

ASJC Scopus subject areas

  • Immunology and Allergy
  • Transplantation
  • Pharmacology (medical)

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