Supplemented αMEM/F12-based medium enables the survival and growth of primary ovarian follicles encapsulated in alginate hydrogels

David Tagler, Yogeshwar Makanji, Nicholas R. Anderson, Teresa K. Woodruff, Lonnie D. Shea*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Hydrogel-encapsulating culture systems for ovarian follicles support the in vitro growth of secondary follicles from various species including mouse, non-primate human, and human; however, the growth of early stage follicles (primary and primordial) has been limited. While encapsulation maintains the structure of early stage follicles, feeder cell populations, such as mouse embryonic fibroblasts (MEFs), are required to stimulate growth and development. Hence, in this report, we investigated feeder-free culture environments for early stage follicle development. Mouse ovarian follicles were encapsulated within alginate hydrogels and cultured in various growth medium formulations. Initial studies employed embryonic stem cell medium formulations as a tool to identify factors that influence the survival, growth, and meiotic competence of early stage follicles. The medium formulation that maximized survival and growth was identified as αMEM/F12 supplemented with fetuin, insulin, transferrin, selenium, and follicle stimulating hormone (FSH). This medium stimulated the growth of late primary (average initial diameter of 80μm) and early secondary (average initial diameter of 90μm) follicles, which developed antral cavities and increased to terminal diameters exceeding 300μm in 14 days. Survival ranged from 18% for 80μm follicles to 36% for 90μm follicles. Furthermore, 80% of the oocytes from surviving follicles with an initial diameter of 90-100μm underwent germinal vesicle breakdown (GVBD), and the percentage of metaphase II (MII) eggs was 50%. Follicle/oocyte growth and GVBD/MII rates were not significantly different from MEF co-culture. Survival was reduced relative to MEF co-culture, yet substantially increased relative to the control medium that had been previously used for secondary follicles. Continued development of culture medium could enable mechanistic studies of early stage folliculogenesis and emerging strategies for fertility preservation.

Original languageEnglish (US)
Pages (from-to)3258-3268
Number of pages11
JournalBiotechnology and Bioengineering
Volume110
Issue number12
DOIs
StatePublished - Dec 1 2013

Keywords

  • Alginate hydrogels
  • Biomaterials
  • Co-culture
  • Culture medium
  • Ovarian follicle development
  • Tissue engineering

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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