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Published on: April 1, 2014
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Decellularization of placentas: establishing a protocol.
L C P C Leonel1, C M F C Miranda1, T M Coelho2
1Setor de Anatomia, Departamento de Cirurgia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo, SP, Brasil.
Summary
Researchers optimized decellularization of canine placentas for tissue engineering. Protocol I better preserved extracellular matrix structure, while Protocol II more effectively removed cells and DNA using sodium dodecyl sulfate (SDS).
Area of Science:
- Biomaterials Science
- Tissue Engineering
- Veterinary Medicine
Background:
- Biological biomaterials for tissue engineering are often derived from decellularized tissues and organs.
- The extracellular matrix (ECM) must remain acellular while preserving its structural and protein integrity.
- Canine placentas offer a readily available source of ECM, but established decellularization protocols are lacking.
Purpose of the Study:
- To establish an effective decellularization method for both maternal and fetal portions of canine placentas.
- To compare the efficacy of different detergents, freezing temperatures, and perfusion techniques.
- To evaluate two distinct decellularization protocols for their impact on ECM preservation and cell removal.
Main Methods:
- Ten preliminary tests evaluated various decellularization parameters.
- Two optimized protocols were selected for detailed analysis.
- Histology, scanning electron microscopy, immunofluorescence, and DNA quantification were employed for assessment.
Main Results:
- Sodium dodecyl sulfate (SDS) proved to be the most effective detergent for cell removal.
- Pre-freezing canine placentas increased the required incubation time for decellularization.
- Both perfusion and immersion methods successfully removed cells.
- Protocol I (1% SDS, 5 mM EDTA, 50 mM TRIS, 0.5% antibiotic) better preserved ECM structure.
- Protocol II (1% SDS, 5 mM EDTA, 0.05% trypsin, 0.5% antibiotic) was more efficient in removing cells and DNA.
Conclusions:
- Decellularization of canine placentas is feasible for generating biomaterials.
- A balance exists between ECM structural preservation and complete cell/DNA removal.
- Further refinement of decellularization protocols is necessary for optimal canine placental biomaterial development.
