Evaluation of sheep ovarian tissue cryopreservation with slow freezing or vitrification after chick embryo chorioallantoic membrane transplantation |
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| Affiliation: | 1. Research and Clinical Center for Infertility, Yazd Reproductive Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran;2. Rafsanjan University of Medical Sciences, Rafsanjan, Iran;1. Laboratory on Animal Germplasm Conservation, Universidade Federal Rural do Semi-Árido – UFERSA, BR 110, Km 47, Costa e Silva, 59625-900, Mossoró, RN, Brazil;2. Laboratory of Manipulation of Oocytes and Preantral Follicles, Universidade Estadual do Ceará – UECE, Paranjana Ave, 1700, Itaperi, 60740-000, Fortaleza, CE, Brazil;3. Laboratory of Wild Animal Biology and Medicine, Faculty of Veterinary Medicine, Federal University of Pará, Belém, Pará, Brazil;4. Schothorst Feed Research, the Netherlands;1. Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, P.O. Box14115-111, Tehran, Iran;2. Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran;1. Department of Animal Science, Food and Nutrition, Southern Illinois University, Carbondale, lllinois, USA;2. Department of Surgery and Obstetrics, College of Veterinary Medicine, University of Baghdad, Baghdad, Iraq;3. Department of Animal and Dairy Sciences, Mississippi State University, Mississippi State, MS, USA;1. Faculty of Veterinary Medicine, Laboratory of Manipulation of Oocytes and Preantral Follicles (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil;2. Institute of Health Sciences, University of International Integration Lusophone African-Brazilian, Acarape, CE, Brazil;3. Federal University of Uberlândia, MG, Brazil;4. Division of Reproductive and Developmental Sciences, Oregon National Primate Research Center, Beaverton, OR, USA;1. Agriculture and Agri-Food Canada, Saskatoon Research and Development Center, Canadian Animal Genetic Resource Program, Saskatoon, SK S7N OX2, Canada;2. Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, SK S7N 5B4, Canada;3. Vaccination and Infectious Disease Organization, University of Saskatchewan, Saskatoon S7N 5E3, Canada |
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| Abstract: | The aim of our investigations was to compare the effectiveness of two methods for cryopreservation of sheep ovarian tissue, slow freezing and vitrification. The quality of cryopreserved tissues was evaluated after 5 days of thawing and chorioallantoic membrane (CAM) transplantation. Follicular structure, stromal integrity and neovascularization were assessed. The areas of fibrosis and necrosis were measured using MICROVISIBLE software, and proliferation was assessed with Ki-67 immunostaning. After 5 days of culture, the proportion of primordial follicles decreased, whereas the primary and intermediary follicles increased insignificantly (p > .05). Only necrosis in the vitrified culture group increased significantly (p < .05). It was established also that 5 days CAM culture was not suitable methodology for detection of folliculogenesis. Follicular quality decreased after culture, but was better in fresh and slow frozen tissues than after vitrification (p < .05). Cellular proliferative activity fell, but it preserved to some extent in all groups. In conclusion, follicles was preserved better in grafted tissue after slow freezing than vitrification and stroma was more susceptible to ischemia in vitrified rather than conventional freezing in this view. Vitrification may not be a suitable alternative to the slow freezing. |
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| Keywords: | Ovarian tissue Slow freezing Vitrification Chorio-allantoic membrane Transplantation |
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