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

• Published in: Journal of Kerman University of Medical Sciences Vol. 27; no. 4; pp. 318 - 328
• Main Authors: Ali Delbari, Maryam Nazm Bojnordi, Sina Mojaverrostami, Hatef Ghasemi Hamidabadi, Zahra Bagheri‑Hosseinabadi, Nourollah Rezaei
• Format: Journal Article
• Language: English
• Published: Kerman University of Medical Sciences 01-07-2020
• Subjects: conditioned medium; embryoid bodies; embryonic stem cells; in vitro maturation
• ISSN: 2008-2843
• DOI: 10.22062/jkmu.2020.91018

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 12 days (first 7 days EBs were cultured in in vitro maturation diluted in Granulose Cell- Conditioned Medium and Follicular Fluid [1:1:1] followed by 5 days of culture in in vitro maturation diluted in uterine condition medium [1:1] ). Results:According tothe MTT test, the viability rate increased in the experimental group compared to the control EBs cultured alone. Expression of Oct4, 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 (ZP1, Figα and GDF9). 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.