Morphology and function of cryopreserved whole ovine ovaries after heterotopic autotransplantation
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BioMed Central
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Morphology and function of cryopreserved whole ovine ovaries after heterotopic autotransplantation Anna T Grazul-Bilska1, Jashoman Banerjee2, Ilker Yazici2, Ewa Borowczyk1, Jerzy J Bilski1, Rakesh K Sharma2, Maria Siemionow3 and Tommaso Falcone*2 Address: 1Department of Animal Sciences, North Dakota State University, Fargo, ND, USA, 2Obstetrics, Gynecology and Women's Health Institute, Cleveland Clinic, Cleveland, OH, USA and 3Department of Plastic Surgery, Cleveland Clinic, Cleveland, OH, USA Email: Anna T Grazul-Bilska - [email protected]; Jashoman Banerjee - [email protected]; Ilker Yazici - [email protected]; Ewa Borowczyk - [email protected]; Jerzy J Bilski - [email protected]; Rakesh K Sharma - [email protected]; Maria Siemionow - [email protected]; Tommaso Falcone* - [email protected] * Corresponding author
Published: 11 April 2008 Reproductive Biology and Endocrinology 2008, 6:16
doi:10.1186/1477-7827-6-16
Received: 4 January 2008 Accepted: 11 April 2008
This article is available from: http://www.rbej.com/content/6/1/16 © 2008 Grazul-Bilska 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: The objective of this study was to perform complex characterization of cryopreserved and then autotransplanted ovaries including determination of the ability to respond to in vivo follicle stimulating hormone (FSH)-treatment, fertilizability of retrieved oocytes, and morphology, vascularization, cellular proliferation and apoptosis in sheep. Methods: Mature crossbred ewes were divided into two groups; an intact (control) group (n = 4), and autotransplanted group (n = 4) in which oophorectomy was performed laparoscopically and ovaries with intact vascular pedicles frozen, thawed and transplanted back into the same animal at a different site. Approximately five months after autotransplantation, estrus was synchronized, ewes were treated with FSH, and ovaries were collected. For all ovaries, number of visible follicles was determined, and collected cumulus oocyte complexes (COC) were matured and fertilized in vitro. Remaining ovarian tissues were fixed for evaluation of morphology, expression of factor VIII (marker of endothelial cells), vascular endothelial growth factor (VEGF; expressed by pericytes and smooth muscle cells), and smooth muscle cell actin (SMCA; marker of pericytes and smooth muscle cells), and cellular proliferation and apoptosis. Two fully functional ovaries were collected from each control ewe (total 8 ovaries). Results: Out of eight autotransplanted ovaries, a total of two ovaries with developing follicles were found. Control ewes had 10.6 +/- 2.7 follicles/ovary, oocytes were in vitro fertilized and developed to the blastocyst stage. One autotransplanted ewe had 4 vi
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