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Effects of different sucrose concentrations on vitrified porcine preantral follicles: Qualitative and quantitative analysis |
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| Institution: | 1. Universidad de Buenos Aires, Facultad de Ciencias Veterinarias, Cátedra de Física Biológica, Instituto de Tecnología e Investigación en Reproducción Animal, Argentina;2. Universidad de Buenos Aires, Facultad de Ciencias Veterinarias, Cátedra de Histología y Embriología, Instituto de Tecnología e Investigación en Reproducción Animal, Argentina;1. School of Biological Sciences, Flinders University, GPO Box 2100, Adelaide, SA 5001, Australia;2. Aquatic Sciences, South Australian Research and Development Institute, 2 Hamra Avenue, West Beach, SA 5024, Australia;3. Department of Fisheries Biology and Genetics, Hajee Mohammad Danesh Science and Technology University, Dinajpur 5200, Bangladesh;1. Yunnan Provincial Engineering Laboratory of Animal Genetic Resource Conservation and Germplasm Enhancement, Yunnan Animal Science and Veterinary Institute, Kunming, Yunnan 650224, People''s Republic of China;2. College of Animal Science and Technology, Yunnan Agricultural University, Kunming, Yunnan 650201, People''s Republic of China;1. Animal Breeding and Reproduction Research Division, NARO Institute of Livestock and Grassland Science, Tsukuba, Japan;2. Embryo Technology and Stem Cell Research Center, Suranaree University of Technology, Nakhon Ratchasima, Thailand;3. Food and Fertilizer Technology Center, Taipei, Taiwan;4. Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul, Korea;1. Departamento de Ciencias de la Salud, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, 09340 DF, Mexico;2. Doctorado en Ciencias Biológicas y de la Salud, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, 09340 DF, Mexico;3. Departamento de Biología de la Reproducción, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, 09340 DF, Mexico;4. Departamento de Medicina y Cirugía Animal, Universidad de Murcia, Espinardo 30100, Spain;1. Department of Surgery, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA;2. Department of Urology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA;3. Department of Obstetrics and Gynecology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA;4. Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA;5. Shriners Hospital for Children, Boston, MA 02114, USA |
| Abstract: | The aim of the present study was to perform a qualitative and quantitative analysis of the effect of different sucrose concentrations combined with ethylene glycol in the preservation of vitrified porcine preantral follicles. Fragments of ovarian cortex were vitrified in cryotubes containing 200 μl of the vitrification solution (30% Ethylene Glycol; 20% Fetal Bovine Serum; 0 M–0.25 M – 0.75 M or 1 M sucrose) and stored in liquid nitrogen for a week. Histological analysis showed that after vitrification the number of normal follicles decreased compared to the fresh tissue (control). The percentage of normal primordial follicles was sucrose dose dependent. The percentage of normal primary follicles was similar in 0 M or 0.25 M sucrose, while higher concentrations (0.75 M and 1 M) increased significantly the percentage of abnormal follicles (p < 0.05). Morphometric analysis showed a statistically significant reduction in the total area of primordial follicles with 0.75 M sucrose and a significant increase in the cytoplasmic area of primordial follicles with 0 M sucrose (p < 0.05). The qualitative and the quantitative analysis appear to be a complementary tool when choosing a vitrification protocol. For our cryopreservation system - vitrification of ovarian cortex slices in cryotubes-the best vitrification medium was TCM 199-Hepes with 30% de ethylene glycol, 20% of Fetal Bovine Serum and 0 or 0.25 M sucrose. The present study shows that the use of high sucrose concentrations in the vitrification solution has a deleterious effect on the preservation of porcine preantral follicles contained in ovarian tissue. Consequently, its use at 0.75 M or 1 M wouldn't be recommended. |
| Keywords: | Porcine Ovarian tissue Preantral follicles Vitrification Cryopreservation CPA"} {"#name":"keyword" "$":{"id":"kwrd0040"} "$$":[{"#name":"text" "_":"Cryoprotective Agent PAF"} {"#name":"keyword" "$":{"id":"kwrd0050"} "$$":[{"#name":"text" "_":"Preantral Follicles BS"} {"#name":"keyword" "$":{"id":"kwrd0060"} "$$":[{"#name":"text" "_":"Base Solution NR"} {"#name":"keyword" "$":{"id":"kwrd0070"} "$$":[{"#name":"text" "_":"Nuclear Roundness OR"} {"#name":"keyword" "$":{"id":"kwrd0080"} "$$":[{"#name":"text" "_":"Oocyte Roundness NA"} {"#name":"keyword" "$":{"id":"kwrd0090"} "$$":[{"#name":"text" "_":"Nuclear Area CA"} {"#name":"keyword" "$":{"id":"kwrd0100"} "$$":[{"#name":"text" "_":"Cytoplasmic Area TA"} {"#name":"keyword" "$":{"id":"kwrd0110"} "$$":[{"#name":"text" "_":"Total Area |
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