Effects of retinoic acid and ovarian somatic cell co-culturing on oocyte differentiation from oocyte-like cells

Bahmanpour, S.; Zareifard, N.; Talaei-Khozani, T.; Esmaeilpour, T.
June 2014
Iranian Journal of Reproductive Medicine;Jun2014 Supplement, Vol. 12, p95
Academic Journal
Introduction: Retinoic acid not only induces meiotic entry in germ cells but also can stimulate proliferation and prevent apoptosis of migratory and postmigratory germ cells. Moreover, growth and development of primary oocytes is dependent on cellular interactions with ovarian somatic cells and production of factors along with changes in oocyte gene expression. This study was designed to determine whether ovarian somatic cell co-culturing and RA can affect differentiation of embryonic stem (ES) cells to oocyte. Materials and Methods: Male mouse ES cells were cultured asembryoid body (EB) for 5 days. Then, each group was co-cultured with ovarian somatic cells in presence or absence of retinoic acid (+RA or -RA) for additional 14 days and cells were assessed for expression of Stra8, SCP3 and Oct4 by quantitative real-time RT-PCR and confirmed for expression of Oct4 by immunocytochemistry. Results: Round cells, ∼15-25 μm, appeard after 3 days of culture on ovarian somatic cells which some of them reached up to 80 μm in size gradually. The gene expression levels of Stra8, SCP3 significantly elevated in the EBs and monolayer in +RA condition compared to -RA condition, in contrast to Oct4 that significantly decreased. Conclusion: This study showed that addition of +RA beside to ovarian somatic cells improved the rate of in vitro differentiation, therefore our approach provides a new strategy for the establishment of oocyte from ES cells.


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