Abstract
In this report, we investigate the fibrin-alginate interpenetrating network (FA-IPN) to provide dynamic cell-responsive mechanical properties, which we apply to the in vitro growth of ovarian follicles. The mechanical properties and polymerization rate of the gels were investigated by rheology, and the fiber structure was imaged by electron microscopy. Using a mouse model, two-layered secondary follicles were encapsulated in FA-IPNs, and growth, morphology, hormone production, fibrin degradation rate and the numbers of competent eggs were assessed. The initial mechanics of the FA-IPN are determined by the composite material, and subsequent degradation of fibrin by the encapsulated cells would produce a material with mechanical properties due to the alginate alone. The rate of meiotically competent oocytes produced by culture in FA-IPN was 82%, which was significantly greater than in alginate alone. This increase in oocyte quality is an important step in identifying 3D culture systems that can provide a fundamental tool to investigate follicle maturation, and may be applied to promote the growth of human follicles, which can be used to provide reproductive options for women facing a cancer diagnosis.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 5476-5485 |
| Number of pages | 10 |
| Journal | Biomaterials |
| Volume | 30 |
| Issue number | 29 |
| DOIs | |
| State | Published - Oct 2009 |
Funding
Androstenedione, 17β-estradiol and progesterone were measured in collected media from 12-day individual follicle culture using commercially available radioimmunoassay kits (androstenedione and 17β-estradiol, Diagnostic Systems Laboratories, Inc., Webster, TX; progesterone, Diagnostic Products Corporation, Los Angeles, CA). The media from the same condition and time point were pulled together, triplicates for each condition. The sensitivities for the androstenedione, estradiol and progesterone assays are 0.1 ng/mL, 10 pg/mL and 0.1 ng/mL, respectively. The hormone assays were performed at the University of Virginia Center for Research in Reproduction Ligand Assay and Analysis Core and was supported by the Eunice Kennedy Shriver NICHD/NIH (SCCPIR) Grant U54-HD28934. The authors would like to thank Baxter International Inc., and more particularly to Drs. Tawil and Hagerman (Baxter BioSurgery, Global R&D, Biotherapeutics and Regenerative Medicine), Dr. Jennifer Hirshfeld-Cytron for helpful scientific discussion, Sarah Kiesewetter and Jennifer Jozefik for their assistance in animal care, Tyler Wellington and PO1 Ovarian Histology Core facility for the assistance in sectioning and staining of the follicles, Stuart Kaltz for his assistance with SEM imaging. This work was supported by U54-HD41857 and P30A: Biomedical Core Oncofertility Consortium Roadmap grant from the NIH:IBIB National Institute of Biomedical Imaging and Bioengineering.
Keywords
- Biodegradation
- Dynamic mechanical properties
- Fibrin-alginate
- IPN (interpenetrating polymer network)
- Ovarian follicle
ASJC Scopus subject areas
- Biophysics
- Bioengineering
- Ceramics and Composites
- Biomaterials
- Mechanics of Materials
