A Novel in vitro Co-culture Systems on Differentiation of Embryonic Stem Cells into Oocyte-like Cells in an in vivo Manner

Background:Differentiation of Embryonic Stem Cells into Oocyte-like cells in vitro is challenging. Successful derivation of oocyte from stem cells can provide an alternative source for curing ovogenesis problems. The current study aims to demonstrate a new protocol with two different types of media for differentiating embryonic stem cells (ESCs) into oocyte-like cells (OLCs). Methods: After culturing mouse ESCs, embryoid bodies (EBs) were generated from ESCs by hanging drop (HD) method. To final differentiation of oocyte-like cells (OLCs), the EBs were cultured in two different types of media for ۱۲ days (first ۷ days EBs were cultured in in vitro maturation diluted in Granulose Cell- Conditioned Medium and Follicular Fluid [۱:۱:۱] followed by ۵ days of culture in in vitro maturation diluted in uterine condition medium [۱:۱] ). Results:According tothe MTT test, the viability rate increased in the experimental group compared to the control EBs cultured alone. Expression of Oct۴, as a pluripotency marker, decreased during the differentiation process of EBs in the experimental group. Co-culturing of EBs with our mentioned protocol increased germ cell markers (Stella and Mvh) and increased Oocyte-specific markers (ZP۱, Figα and GDF۹). Conclusion:Our study introduces a promising in vitro protocol for achieving successful oogenesis through creating interactions of EBs with granulosa cells and uterine condition medium.

کلیدواژه ها:

Embryonic Stem Cells ، Conditioned Medium ، Embryoid Bodies ، in vitro maturation