Tracing ovarian follicles growth in vitro by three-dimensional culture system

Abstract

In female mammals, the basic reproductive unit in the ovaries is the follicle, which is composed by the oocyte and surrounding somatic granulosa cells (GCs). The development of follicles is a highly programed progress and is regulated by both the paracrine and endocrine factors. To culture the follicle in vitro, various approaches were established in different groups including the 2-dimensional culture and 3-dimensional culture. In this study, we improved a three-dimensional Matrigel system to culture the mouse follicles from secondary stage. In this system, the 3D culture guaranteed spherical structures of the follicles, which was utilized to trace the difference of growth dynamics of follicles in mutant and wild type females.

Simulated microgravity reduces quality of ovarian follicles and oocytes by disrupting communications of follicle cells

“…To investigate the influences of microgravity on the development of ovarian follicles and oocytes in mice, we modified a three-dimensional follicle culture system in a RCCS to support mouse ovarian follicle development under the SMG condition 16,23 . This was achieved by seeding single follicle (about 200 μm diameter) into liquid Matrigel droplets, which were then transformed to a solid gel in the culture to support follicle growth.…”

“…4). To observe the growth of follicles, the follicles were cultured in Matrigel (BD, 354234) culture system as previously reported 23,[55][56][57] . The culture medium consisted of 10 mL MEMα (Gibco, 32-571-036), 2.1 mg/mL NaHCO 3 , 5% FBS (Gibco, 10-099-141), 100 IU/mL penicillin-streptomycin (15140122, Invitrogen), 1% ITS (Sigma-Aldrich, 13146) and 10 ng/mL FSH (ovine Follicle Stimulating Hormone, NHPP).…”