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Comparative characterization of decellularized renal scaffolds for tissue engineering
Iris Fischer1, Michael Westphal1, Bella Rossbach1
1Berlin-Brandenburg Center for Regenerative Therapies, Charité-Universitätsmedizin Berlin, Augustenburger Platz 1, D-13353 Berlin, Germany.
Biomedical Materials (Bristol, England)
|April 12, 2017
Summary
This study standardized kidney decellularization methods, finding SDS best for structure and SDC best for cell function. A scoring system aids comparison of extracellular matrix (ECM) preservation strategies.
Area of Science:
- Biomaterials Science
- Tissue Engineering
- Regenerative Medicine
Background:
- Native extracellular matrix (ECM) is crucial for tissue engineering scaffolds, providing architectural and biochemical cues.
- Decellularized ECM (dECM) retains native cues, promoting cell arrangement and function for tissue regeneration.
- Inconsistent decellularization and assessment methods hinder comparability of kidney dECM strategies.
Purpose of the Study:
- To standardize and compare decellularization protocols for porcine kidney ECM using different detergents and temperatures.
- To evaluate the impact of decellularization on ECM structure, composition, and cell attachment/viability.
- To develop a scoring system for comparing decellularization strategies and guiding future research.
Main Methods:
- Porcine kidney tissue was decellularized using a standardized immersion protocol with Triton X-100, SDS, and SDC at varying temperatures.
- Decellularization efficacy, ECM structural integrity, glycosaminoglycan (GAG), collagen, and cytokine content were analyzed.
- Attachment and viability of human induced pluripotent stem cell-derived renal precursor cells on dECM were assessed.
Main Results:
- Structural ECM preservation was temperature-dependent; GAG and collagen were best maintained with 1% SDS at 4°C.
- Cytokine preservation was superior with 1% SDC at 4°C, correlating with better cell attachment and viability.
- A novel scoring system revealed SDS at 4°C yielded highest structural/composition scores, while SDC at 4°C showed better cell performance.
Conclusions:
- Decellularization strategy significantly impacts ECM structure, composition, and cell function; SDS and SDC offer distinct advantages.
- The developed scoring system enables standardized comparison of decellularization methods across studies.
- Cell attachment and viability are critical, independent parameters that must be included in all decellularization assessments.