Disclosure statement: No competing financial interests exist.
Supplemented αMEM/F12-based medium enables the survival and growth of primary ovarian follicles encapsulated in alginate hydrogels
Article first published online: 15 JUL 2013
© 2013 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 110, Issue 12, pages 3258–3268, December 2013
How to Cite
Tagler, D., Makanji, Y., Anderson, N. R., Woodruff, T. K. and Shea, L. D. (2013), Supplemented αMEM/F12-based medium enables the survival and growth of primary ovarian follicles encapsulated in alginate hydrogels. Biotechnol. Bioeng., 110: 3258–3268. doi: 10.1002/bit.24986
- Issue published online: 23 OCT 2013
- Article first published online: 15 JUL 2013
- Accepted manuscript online: 25 JUN 2013 06:26AM EST
- Manuscript Accepted: 17 JUN 2013
- Manuscript Revised: 12 JUN 2013
- Manuscript Received: 10 MAY 2013
- National Institutes of Health (NIH). Grant Number: PL1EB008542
- Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD). Grant Numbers: U54HD41857, U54HD076188
- National Health and Medical Research Council (NHMRC) (Australia). Grant Number: GNT1016460
- ovarian follicle development;
- alginate hydrogels;
- culture medium;
- tissue engineering
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 80 µm) and early secondary (average initial diameter of 90 µm) follicles, which developed antral cavities and increased to terminal diameters exceeding 300 µm in 14 days. Survival ranged from 18% for 80 µm follicles to 36% for 90 µm follicles. Furthermore, 80% of the oocytes from surviving follicles with an initial diameter of 90–100 µm 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. Biotechnol. Bioeng. 2013;110: 3258–3268. © 2013 Wiley Periodicals, Inc.