Human Placenta Decellularized Matrix as A Scaffold for Use in Proliferation of Spermatogonial Stem Cells

Background: Extracellular matrix (ECM) of tissues contains various ranges of growth factors, proteins, proteoglycans, hya-luronic acid and others. Decellularization of tissue and produc-tion of tissue engineering scaffolds from ECM components is one of the most reliable strategies in fabrication of scaffolds. Although various protocols have been developed to remove the cells from tissues with minimal degradation of ECM compo-nents, an optimal protocol is still needed to satisfy scientists. In this study, we examined various procedures for decellulariza-tion of placenta and optimized a reliable decellularization pro-tocol.Materials and Methods: Human placenta was obtained from mothers undergoing cesarean after obtaining informed consent. The tissues were treated with various contents of Triton X-100 and sodium dodecyl sulphate (SDS) for 15 or 30 minutes. The decellularized tissues were casted, freezed and freeze dried to fabricate a porous scaffold. The scaffolds were then cross-linked by glutaraldehyde gas. The removal of the cells from tis-sues was determined by H&E and 4′, 6-diamidino-2-phenylin-dole (DAPI) staining, and DNA content assay. Alcian blue, Masson’s trichrome and Orcein staining was used to confirm that ECM remained intact after decellularization process. Mor-phology of porous scaffold was viewed under scanning electron microscopy (SEM). Cell viability and cell adhesion property of scaffold were determined by MTT and SEM.Results: Histological analysis showed that the groups content %0.5 SDS and SDS+ Triton X-100 for 30 minutes were com-pletely decellularized. Decellularization was further confirmed by DAPI staining and DNA content assay. Laminin, collaegen, elastin and glycosaminoglycans remained intact after decellularization process. MTT test showed cell viability changes for %0.5 SDS group. SDS+ Triton X-100 for 30 minutes have no significant difference with control group (P> 0.05).Conclusion: Our study proved a reliable and effective protocol for decellularization of placenta with minimal negative effects on ECM components. The decellularized placenta is suggested as a promising bio-scaffold tissue engineering applications.

کلیدواژه ها:

Decellularized Scaffold ، Placenta ، Extracellular ma-trix