Abstract:

BACKGROUND: Whether umbilical mesenchymal stem cells (UC-MSCs) can be induced into oocytes under in vitro and in vivo microenvironment is very important for female reproductive maintenance and oocyte regeneration.
OBJECTIVE: To further testify the effects of embryonic bodies (EB) under in vitro microenvironment on the differentiation of UC-MSCs into germ cells.
METHODS: UC-MSCs were cultured in hanging drop to form EB, which were co-cultured with human ovarian granulosa cells or mouse ovarian granulosa cells, or were cultured in follicular fluid to induce differentiation of UC-MSCs into primary germ cells in vitro.
RESULTS AND CONCLUSION: The EB were successfully prepared when UC-MSCs were cultured in hanging drop. Flow cytometry results: SSEA-1+ cells accounted for 15.61% after 5 days of EB formation. The results of immunohistochemistry: EB of 5 days were co-cultured with human ovarian granulosa cells or mouse ovarian granulosa cells for 10 days, germ-line markers of synaptonemal complex protein-3 and growth differentiation factor-9 positively expressed, while these two markers did not express in granule cells or EB cultured in follicular fluid. EB can be formed when UC-MSCs are cultured in hanging drop in vitro. Germ-line markers will express in EB when they are co-cultured with human ovarian granulosa cells or mouse ovarian granulosa cells.