title: Promoting extracellular matrix remodeling via ascorbic acid enhances the survival of primary ovarian follicles encapsulated in alginate hydrogels.
abstract: The in vitro growth of ovarian follicles is an emerging technology for fertility preservation. Various strategies support the culture of secondary and multilayer follicles from various species including mice, non-human primate, and human; however, the culture of early stage (primary and primordial) follicles, which are more abundant in the ovary and survive cryopreservation, has been limited. Hydrogel-encapsulating follicle culture systems that employed feeder cells, such as mouse embryonic fibroblasts (MEFs), stimulated the growth of primary follicles (70-80 µm); yet, survival was low and smaller follicles (<70 µm) rapidly lost structure and degenerated. These morphologic changes were associated with a breakdown of the follicular basement membrane; hence, this study investigated ascorbic acid based on its role in extracellular matrix (ECM) deposition/remodeling for other applications. The selection of ascorbic acid was further supported by a microarray analysis that suggested a decrease in mRNA levels of enzymes within the ascorbate pathway between primordial, primary, and secondary follicles. The supplementation of ascorbic acid (50 µg/mL) significantly enhanced the survival of primary follicles (<80 µm) cultured in alginate hydrogels, which coincided with improved structural integrity. Follicles developed antral cavities and increased to diameters exceeding 250 µm. Consistent with improved structural integrity, the gene/protein expression of ECM and cell adhesion molecules was significantly changed. This research supports the notion that modifying the culture environment (medium components) can substantially enhance the survival and growth of early stage follicles.
authors.affiliation: Department of Chemical and Biological Engineering, McCormick School of Engineering, Northwestern University, Technological Institute E-136, 2145 Sheridan Road, Evanston, Illinois, 60208; Institute for BioNanotechnology in Medicine (IBNAM), Northwestern University, 303 E. Superior Street, Suite 11-131, Chicago, Illinois, 60611.
authors.forename: David
authors.initials: D
authors.lastname: Tagler
authors.forename: Yogeshwar
authors.initials: Y
authors.lastname: Makanji
authors.forename: Tao
authors.initials: T
authors.lastname: Tu
authors.forename: Beatriz Peñalver
authors.initials: BP
authors.lastname: Bernabé
authors.forename: Raymond
authors.initials: R
authors.lastname: Lee
authors.forename: Jie
authors.initials: J
authors.lastname: Zhu
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Kniazeva
authors.forename: Jessica E
authors.initials: JE
authors.lastname: Hornick
authors.forename: Teresa K
authors.initials: TK
authors.lastname: Woodruff
authors.forename: Lonnie D
authors.initials: LD
authors.lastname: Shea
mesh_terms: D000464:Alginates / Q000378:metabolism*; D000818:Animals; D001205:Ascorbic Acid / Q000378:metabolism*; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry; D005109:Extracellular Matrix / Q000187:drug effects*; D005260:Female; D020723:Glucuronic Acid / Q000378:metabolism; D006603:Hexuronic Acids / Q000378:metabolism; D020100:Hydrogels / Q000378:metabolism*; D051379:Mice; D006080:Ovarian Follicle / Q000502:physiology*
chemical_list: D000464:Alginates; D003470:Culture Media; D006603:Hexuronic Acids; D020100:Hydrogels; D020723:Glucuronic Acid; D001205:Ascorbic Acid
keywords: alginate hydrogel; ascorbic acid; biomaterial; extracellular matrix; ovarian follicle development; primary follicle; regenerative medicine; tissue engineering