Abstract
Mesenchymal stem cells (MSCs) secrete a variety of cytokines and growth factors in addition to self-renewal and multiple forms of differentiation. Some of these secreted bioactive factors could improve meiotic maturation in vitro and subsequent embryo developmental potential. The aim of the present study was to determine whether in vitro maturation (IVM) of mouse oocyte with or without cumulus cells could be improved by contact with conditioned medium (CM) of MSCs as well as the efficiency of CM to support follicular growth and oocyte maturation in the ovarian organ of mice cultured on soft agar. The developmental potential of matured oocyte was assessed by blastocyst formation after in vitro fertilization (IVF). Germinal vesicle stage oocytes with or without cumulus cells were subjected to IVM in either CM, Dulbecco's modified Eagle's medium (DMEM), alpha-minimum essential medium (alpha-MEM) or human tubal fluid (HTF). Approximately 120 oocytes were studied for each medium. CM produced a higher maturation rate (91.2%) than DMEM (54.7%), alpha-MEM (63.5%) and HTF (27.1%). Moreover, CM improved embryo development to blastocyst stage significantly more than DMEM and HTF (85 vs 7% and 41.7%, respectively) but there was no significant difference compared with alpha-MEM (85 vs 80.3%). The behavior of cortical granules of IVM oocytes cultured in CM revealed cytoplasmic maturation. Moreover, CM also supported preantral follicles growth well in organotypic culture on soft agar resulting in the maturation of 60% of them to developmentally competent oocytes. The production of estrogen progressively increased approximately 1-fold every other day during organ culture, while a dramatic 10-fold increase in progesterone was observed 17 h after human chorionic gonadotropin stimulus at the end of culture. Thus, CM is an effective medium for preantral follicle growth, oocyte maturation, and sequential embryo development.
Highlights
Technologies to stimulate oocyte growth and development in culture can provide valuable experimental tools for studies of the mechanisms governing oocyte development as well as support practical clinical, agricultural, and zoological applications
Since it is known that cytokines and growth factors stimulate meiotic progress and the processes associated with in vitro maturation (IVM), the aim of the present study was to determine whether IVM with cumulus cells in mice could be improved by conditioned medium (CM), and the efficiency of CM for in vitro follicular growth of organotypic culture on soft agar
We investigated the ability of CM to support in vitro www.bjournal.com.br follicular growth in organotypic culture and IVM of germinal vesicle (GV) oocytes and subsequent embryo development
Summary
Technologies to stimulate oocyte growth and development in culture can provide valuable experimental tools for studies of the mechanisms governing oocyte development as well as support practical clinical, agricultural, and zoological applications. Many attempts have been made to grow immature oocytes in vitro [1,2,3]. It has been established that the culture conditions used for in vitro maturation (IVM), which include different media, type and hormone content, as well as the presence of cumulus cells, can significantly influence maturation rates and sub-. It is known that oocytes need to undergo cytoplasmic maturation as well as nuclear maturation to become able to support successful fertilization and embryo development [9,10]. Nuclear maturation can be identified with germinal vesicle breakdown (GVBD) and polar body extrusion, while cytoplasmic maturation is hard to identify due to the absence of a direct marker
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