LAUSR

Three-dimensional (3D) culture provides a biomimicry of the naive microenvironment that can support cell proliferation, differentiation and regeneration. Some growth factors, such as epidermal growth factor (EGF), facilitate normal meiosis during oocyte maturation in vivo. In this study, a scaffold-based 3D co-culture system, using purified alginate was applied to induce oocyte differentiation from mouse embryonic stem cells (mESCs). Mouse embryonic stem cells were induced to differentiate into oocyte-like cells using embryoid body (EB) protocol in the two-dimensional (2D) or 3D microenvironment in-vitro. To increase the efficiency of the oocyte-like cells differentiation from mESCs, we employed a co-culture system using ovarian granulosa cells in the presence or absence of epidermal growth factor (+EGF or -EGF) for 14 days and then the cells were assessed for germ cell differentiation, meiotic progression and oocyte maturation markers. The cultures exposed to EGF in the alginate-based 3D microenvironment, showed the highest level of premeiotic (Oct4, Mvh), meiotic (Scp1, Scp3, Stra8, Rec8) and oocyte maturation (Gdf9, Cx37, Zp2) marker genes (P<0.05) in comparison to other groups. According to the gene expression patterns, we can conclude that alginate-based 3D co-culture system provided a highly efficient protocol for oocyte-like cell differentiation from mESCs. The data showed that this culture system along with EGF improved the rate of in vitro oocyte-like cell differentiation. This article is protected by copyright. All rights reserved.