Comparing various protocols of human and bovine ovarian tissue decellularization to prepare extracellular matrix-alginate scaffold for better follicle development in vitro

Hossein Nikniaz,Neda Daei-Farshbaf,Seyed Behnamedin Jameie,Mohammad Nouri,Zahra Zandieh,Mazaher Gholipourmalekabadi,Reza Aflatoonian

doi:10.1186/s12896-020-00658-3

Abstract

BackgroundNowadays, the number of cancer survivors is significantly increasing as a result of efficient chemo/radio therapeutic treatments. Female cancer survivors may suffer from decreased fertility. In this regard, different fertility preservation techniques were developed. Artificial ovary is one of these methods suggested by several scientific groups. Decellularized ovarian cortex has been introduced as a scaffold in the field of human fertility preservation. This study was carried out to compare decellularization of the ovarian scaffold by various protocols and evaluate the follicle survival in extracellular matrix (ECM)-alginate scaffold.ResultsThe micrographs of H&E and DAPI staining confirmed successful decellularization of the ovarian cortex in all experimental groups, but residual DNA content in SDS-Triton group was significantly higher than other groups (P < 0.05). SEM images demonstrated that complex fiber network and porosity structure were maintained in all groups. Furthermore, elastin and collagen fibers were observed in all groups after decellularization process. MTT test revealed higher cytobiocompatibility of the SDS-Triton-Ammonium and SDS-Triton decellularized scaffolds compared with SDS groups. Compared to the transferred follicles into the sodium alginate (81%), 85.9% of the transferred follicles into the decellularized scaffold were viable after 7 days of cultivation (P = 0.04).ConclusionAlthough all the decellularization procedures was effective in removal of cells from ovarian cortex, SDS-Triton-Ammonium group showed less residual DNA content with higher cytobiocompatibility for follicles when compared with other groups. In addition, the scaffold made from ovarian tissues decellularized using SDS-Triton-Ammonium and sodium alginate is suggested as a potential 3D substrate for in vitro culture of follicles for fertility preservation.

Highlights

Nowadays, the number of cancer survivors is significantly increasing as a result of efficient chemo/radio therapeutic treatments [1]
Decellularization of bovine ovarian cortex The hematoxylin and eosin (H&E) staining results showed that ovarian tissues were completely decellularized in all groups (Fig. 1a-e)
Staining of elastic fibers and collagen proteins with orcein (Fig. 1 K-O) and trichrome

Summary

Introduction

The number of cancer survivors is significantly increasing as a result of efficient chemo/ radio therapeutic treatments. Female cancer survivors may suffer from decreased fertility In this regard, different fertility preservation techniques were developed. Scientific groups attempt to find ideal scaffold to support follicle development [10] To reach this aim, different kinds of natural and synthetic matrixes were examined. SDS is the ionic surfactant and a prevalent agent used in the decellularization process and successfully eliminates cells’ nuclear materials, it can be harmful to the structural of ECM and signaling proteins [15]. It is a cytotoxic agent and irremovable due to its ionic nature [11]. Ammonium retains glycosaminoglycans (GAGs) and viscoelasticity of the decellularized tissue by counteracting the negative charge of collagen [17]

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Discussion

Conclusion