Optimization of a decellularization protocol for the production of scaffolds from murine placentas

Placentas as organs of reproduction that are well vascularized and that possessed an extensive extracellular matrix to preserve the inner structure are promising for building biological scaffolds. To establish an effective decellularization protocol for murine placentas we used 40 mice placentas from C57BL/6 of 17.5-20 days of gestation and 20 placentas of Wistar rats of 14.5 days, tested for 4 protocols and controls were performed by histology and immunohistochemistry for collagen I, III and IV, fibronectin and laminin. The protocols included washing with PBS (Phosphate-Buffered Saline) 1X + EDTA (Ethylenediaminetetraacetic acid) + ATB (Antibiotics Penicillin/Streptomycin) 0.5%: (P1) 0.25% SDS (Sodium Dodecyl Sulfate) for 72 hours; (P2) 0.3% SDS for 72 hours; (P3) 0:25%, 0.5%, 1.0% SDS each for 24 hours; (P4) 0.001% 0.01% and 1.0% SDS for 24 hours each. Finally, incubation with Triton X-100 1% followed for 48 hours in all groups. For mice, protocol 1 resulted as the best, whereas decellularization in rats was successful by protocol 2. Controls by histology revealed the absence of nuclei and cells in these samples. Immunohistochemistry showed the presence of collagens, fibronectin and laminin in the extracellular matrices. Especially in mice, they were mainly found in the labyrinth zone in contrast to the junctional zone towards the decidua. Ongoing research concentrates on the biological meaning of this finding. In conclusion, we found adequate protocols for the decellularization of mice and rat placentas in order to build biological scaffolds for further applications.