Calcium alginate microencapsulation of ovarian follicles impacts FSH delivery and follicle morphology
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BioMed Central
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Calcium alginate microencapsulation of ovarian follicles impacts FSH delivery and follicle morphology Matthew Heise1, Richard Koepsel1, Alan J Russell1 and Elizabeth A McGee*1,2,3 Address: 1McGowan Institute for Regenerative Medicine, University of Pittsburgh, 100 Technology Dr. Suite 200, Pittsburgh, PA 15219, USA, 2Magee-Womens Research Institute, 204 Craft Ave, Pittsburgh PA, 15213, USA and 3Department of Obstetrics, Gynecology and Reproductive Medicine, University of Pittsburgh, Pittsburgh PA 15213, USA Email: Matthew Heise - [email protected]; Richard Koepsel - [email protected]; Alan J Russell - [email protected]; Elizabeth A McGee* - [email protected] * Corresponding author
Published: 14 September 2005 Reproductive Biology and Endocrinology 2005, 3:47
doi:10.1186/1477-7827-3-47
Received: 19 April 2005 Accepted: 14 September 2005
This article is available from: http://www.rbej.com/content/3/1/47 © 2005 Heise et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract Background: We have previously shown that suspension culture prevents follicle flattening and maintains three-dimensional follicle architecture better than culture on flat plates. However, many of the follicles cultured in suspension do eventually rupture, as basement membrane integrity is lost and the three-dimensional structure of the follicle is altered. Therefore, the objective of this study is to support three-dimensional follicle architecture during in vitro growth of ovarian follicles through encapsulation in calcium alginate, while maintaining responsiveness to FSH stimulation. Methods: Preantral follicles (150 – 160 micrometers in diameter) were isolated from the ovaries of juvenile rats and grown in culture tubes or encapsulated in calcium alginate and grown in culture tubes. Previous studies revealed that follicles maintained structural integrity but did not grow as well when encapsulated in calcium alginate. In these studies, we evaluated the effect of calcium alginate on FSH-stimulated follicle growth, survival, and morphology in suspension culture. Follicles were grown under 5 culture conditions: 1) not encapsulated; with FSH in the medium, 2) encapsulated in the absence of FSH, grown in medium without FSH, 3) encapsulated with calcium alginate containing FSH but grown in medium without FSH, 4) encapsulated without FSH but grown in medium containing FSH and 5) encapsulated with calcium alginate containing FSH and in medium containing FSH. To assess growth rates, follicles were cultured for 72 hours and analyzed for follicle size increase and DNA content. Survival analysis for encapsulated and unencapsulated follicles was performed by constructing a Kaplan Meier survival curve of daily observations of intact follicle survival. Three-dimensi
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