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    Comparison of cryoprotective effects of iodixanol, trehalose and cysteamine on ram semen
    (Elsevier, 2013) Cirit, Umut; Bagis, Haydar; Demir, Kamber; Agca, Cansu; Pabuccuoglu, Serhat; Varisli, Omer; Clifford-Rathert, Charlotte
    This study was conducted to improve cryosurvival of electroejaculated (EE) ram semen in the presence of iodixanol (OptiPrep (TM)), trehalose or cysteamine. A tris-based extender was used to prepare 12 extenders containing OptiPrep (TM) (Op), trehalose (Tr) or cysteamine (Cy) alone, or different combinations of these compounds. Extenders were designated as follows: Tris (control), Op1.25 (1.25% Op, v/v), Op2.5 (2.5% Op, v/v), Op5 (5% Op, v/v), Tr50 (50 mM Tr), Tr100 (100 mM Tr), Cy (5 mM Cy), OpTr (2.5% Op and 100 mM Tr), OpCy (2.5% Op and 5 mM Cy), TrCy (100 mM Tr and 5 mM Cy), OpTrCy1 (2.5% Op, 100 mM Tr and 5 mM Cy) and OpTrCy2 (1.25% Op, 50 mM Tr and 2.5 mM Cy). A two-step dilution was used and glycerol was added at 5 degrees C in the second step. Diluted samples were equilibrated for 1 h, loaded in 0.25 mL straws and frozen in a programmable freezing machine. Supplementation of 5% OptiPrep (TM) significantly protected post-thaw progressive motility, membrane integrity, acrosomal integrity and morphological damages. Trehalose supplementation protected membrane integrity of ram sperm; however, it did not help post-thaw motility and morphology. Supplementation of 5 mM cysteamine had detrimental effect on cryosurvival of EE ram semen. These results demonstrate that the supplementation of iodixanol increases the cryosurvival of EE ram semen in a dose-dependent manner. (c) 2013 Elsevier B.V. All rights reserved.
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    Development of starch based mucoadhesive vaginal drug delivery systems for application in veterinary medicine
    (Elsevier Sci Ltd, 2016) Gok, Mehmet Koray; Ozgumus, Saadet; Demir, Kamber; Cirit, Umut; Pabuccuoglu, Serhat; Cevher, Erdal; Ozsoy, Yildiz
    The aim of this study was to prepare and evaluate the mucoadhesive, biocompatible and biodegradable progesterone containing vaginal tablets based on modified starch copolymers for the estrus synchronization of ewes. Starch-graft-poly( acrylic acid) copolymers (S-g-PAA) were synthesized and characterized. The vaginal tablets were fabricated with S-g-PAA and their equilibrium swelling degree (Qe) and matrix erosion (ME%) were determined in lactate buffer solution. In vitro, mucoadhesive properties of the tablets were investigated by using ewe vaginal mucosa and in vivo residence time were also investigated. In vitro and in vivo progesterone release profiles from the tablets were compared with two commercial products. Tablet formulation containing wheat starch based grafted copolymer (WS-g-PAA)(gc) indicated promising results and might be convenient as an alternative product to the commercial products in veterinary medicine. (C) 2015 Elsevier Ltd. All rights reserved.
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    Effect of different transport temperatures on in vitro maturation of oocytes collected from frozen-thawed sheep ovaries
    (Tubitak Scientific & Technological Research Council Turkey, 2013) Ozdas, Ozen Banu; Baran, Alper; Tas, Muzaffer; Cirit, Umut; Demir, Kamber; Bacinoglu, Suleyman; Pabuccuoglu, Serhat
    The aim of this study was to determine the effects of 2 different transport temperatures on the in vitro maturation of oocytes collected from frozen-thawed sheep ovaries. Sheep ovaries were transferred into saline at temperatures of 4 degrees C and 32 degrees C. After the 2 experimental groups (A: fresh cortex, B: frozen-thawed cortex) were formed, each group was divided into 2 subgroups (group A1: 4 degrees C, group A2: 32 degrees C [control]; group B1: 4 degrees C, group B2: 32 degrees C). The cortexes were dissected into slices 1-3 mm thick and pieces of 0.5 cm(2). For groups B1 and B2, 1-2 cortex pieces were placed in cryogenic vials containing 1 mL of freezing medium modified with Earle's salts (TCM-199) and supplemented with 10% fetal calf serum (FCS) (FCS + 2.5 M ethylene glycol + 0.1 M sucrose). The vials were then cooled to 7 degrees C at 2 degrees C/min and held at 7 degrees C for 10 min for manual seeding. The temperature was then lowered by 0.3 degrees C/min to -35 degrees C and thereafter by -10 degrees C/min to -75 degrees C. Vials were plunged into -196 degrees C liquid nitrogen and stored. Cortexes were thawed at 37 degrees C. Collected oocytes were matured in their own groups in 700 mu L of TCM-199 (supplemented with luteinizing hormone, follicle-stimulating hormone, pyruvate, and FCS) for 23 h in a gas mixture of 5% CO2, 5% O-2, and 90% N-2 at 38.8 degrees C. After maturation, oocytes were fixed in acetic acid and ethyl alcohol (1: 3) for 48 h. Oocytes were stained with aceto-orcein and then examined. At the end of the study, maturation rates for reaching metaphase I (MI) were similar in all groups (group A1: 30.76%, group A2: 38.09%, group B1: 30.65%, and group B2: 33.33%). The rates at which metaphase II (MII) was reached were 18.58%, 34.69%, 7.25%, and 6.48%, respectively. The best development was seen in group A2 (P < 0.001). Sheep oocytes obtained from fresh and frozen-thawed cortexes reached the MII stage if transported at 4 degrees C.
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    Effect of transport and storage temperature of ovaries on in vitro maturation of bitch oocytes
    (Elsevier Science Bv, 2006) Tas, Muzaffer; Evecen, Mithat; Ozdas, Ozen Banu; Cirit, Umut; Demir, Kamber; Birler, Sema; Pabuccuoglu, Serhat
    In this study, the effects of ovary transport and storage temperature on in vitro maturation of bitch oocytes were investigated. Ovaries were collected from 23 mature bitches and one randomly selected ovary of each pair (n = 23 pairs) was transported in physiologic saline at 4 degrees C, while the other one at 35-38 degrees C for 2-4 h. A total of 316 cumulus oocyte complexes (COCs) were obtained from the 4 degrees C group and 301 COCs from the 35-38 degrees C group. All COCs were matured in modified synthetic oviduct fluid (mSOF) supplemented with follicle stimulating hormone (FSH), essential and non-essential amino acids at 38 degrees C in ahumidified 5% CO2, 5% O-2, and 90% N-2 atmosphere for 72 h. At the end of the in vitro maturation period, nuclear maturation of oocytes was classified as germinal vesicle (GV), germinal vesicle breakdown (GVBD), metaphase I (MI), metaphase II (MII), undetermined nuclear maturation (UDNM), and MI + MII. The nuclear maturation rates to MI, MII, and MI + MII stages were 60.44%, 10.75%, and 71.20% in the 4 degrees C group and 37.20%, 7.64%, and 45.85% in the 35-38 degrees C group, respectively. The data demonstrated that oocytes obtained from ovaries transported at 4 degrees C had higher maturation rates than from the ones transported at 35-38 degrees C (p < 0.001). (c) 2005 Published by Elsevier B.V.
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    Effects of estrous cycle stage and transport temperature of ovaries on in vitro maturation of canine oocytes
    (Elsevier Science Bv, 2010) Evecen, Mithat; Cirit, Uemuet; Demir, Kamber; Ozdas, Ozen Banu; Tas, Muzaffer; Birler, Sema; Pabuccuoglu, Serhat
    Unlike other domestic animals, in vitro maturation (IVM) of canine oocytes still has limited success. The present study investigated the effects of estrous cycle stage and transport temperature of ovaries on in vitro maturation of canine oocytes. The donor bitches were categorized into three groups based on stage of estrus cycle: follicular (proestrus or estrous), luteal (diestrus) and anestrus. One ovary of each pair collected from 39 mature bitches was transported in Phosphate Buffer Saline (PBS) at 4 degrees C while the other was transported at 37 degrees C. A total of 1138 Grade I COCs obtained from all ovaries were grouped and matured in modified synthetic oviduct fluid (mSOF) supplemented with follicle stimulating hormone (FSH), luteinizing hormone (LH), essential and non-essential amino acids at 38.5 degrees C in a humidified 5% CO2, 5% O-2, and 90% N-2 atmosphere for 72 h. The nuclear maturation rates were evaluated by aceto-orcein staining. Oocytes harvested from follicular and luteal ovaries have a significantly higher maturation rates (MI+MII) than the oocytes from anestrual ovaries in the 37 degrees C group (p<0.05). However, oocytes harvested from anestrual ovaries transported at 4 degrees C had the highest maturation (MI + MII) rate, and the difference between anestrual and luteal ovary groups was significant (p < 0.05). The oocytes from anestrual ovaries transported at 4 degrees C have significantly higher maturation rates than those transported at 37 degrees C (p < 0.0001). However, the transport temperature (37 or 4 degrees C) did not significantly affect the maturation (MI + MII) rates of oocytes harvested from the luteal (p = 0.61) and follicular (p = 0.48) stage ovaries. It can be concluded from this study that (1) both transport temperature and transport temperature x estrus cycle stage interaction effected the maturation rates, while estrus cycle stage alone did not, and (2) transporting canine ovaries at 4 degrees C can improve in vitro maturation rates in oocytes harvested from anestrous ovaries. (C) 2009 Elsevier B.V. All rights reserved.
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    Evaluation of synergic effects of iodixanol and trehalose on cryosurvival of electroejaculated ram semen
    (Wiley, 2020) Ozmen, Mehmet Ferit; Cirit, Umut; Arici, Ramazan; Demir, Kamber; Kurt, Dogan; Pabuccuoglu, Serhat; Ak, Kemal
    The primary aim of the study was to investigate whether iodixanol and trehalose would have a synergic effect on the cryosurvival of electroejaculated ram semen. Tris-based diluter was used to prepare 9 different extenders by the addition of iodixanol or trehalose alone or varying combinations of these substances. Diluters were prepared as follows: Tris (control), Io5 (5% iodixanol), Tr25 (25 mmol/L trehalose), Tr50 (50 mmol/L trehalose), Tr50 + Io1.25 (50 mmol/L trehalose and 1.25% iodixanol), Tr50 + Io2.5 (50 mmol/L trehalose and 2.5% iodixanol), Tr50 + Io5 (50 mmol/L trehalose and 5% iodixanol), Tr25 + Io5 (25 mmol/L trehalose and 5% iodixanol) and Tr12.5 + Io5 (12.5 mmol/L trehalose and 5% iodixanol). Supplementation of the freezing extender with trehalose or iodixanol alone supported the protection of both morphological and functional integrity of ram spermatozoa and total motility at 1 and 4 hr post-thawing respectively. However, beyond these positive effects, the combination of trehalose (25 mmol/L) and iodixanol (5%) significantly increased post-thaw sperm longevity and motion properties at the end of 4-hr incubation. The results of the study clearly showed that there was positive synergic effect of iodixanol and trehalose on cryosurvival of ram semen.
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    Using cell banks as a tool in conservation programmes of native domestic breeds: the production of the first cloned Anatolian Grey cattle
    (Csiro Publishing, 2011) Arat, Sezen; Caputcu, Arzu T.; Akkoc, Tolga; Pabuccuoglu, Serhat; Sagirkaya, Hakan; Cirit, Umut; Nak, Yavuz
    The aim of this study was to clone native Anatolian Grey cattle by using different donor cell types, such as fibroblast, cartilage and granulosa cells cryopreserved in a gene bank and oocytes aspirated from ovaries of Holstein cows as the recipient cytoplasm source. One male calf from fibroblast, three female calves from granulosa cells and one female calf from cartilage cells were born healthy and at normal birthweights. No calves were lost after birth. The results demonstrated that the cloned calves had the same microsatellite alleles at 11 loci as their nuclear donors. However, the mtDNAs of the five Anatolian Grey cloned calves had different haplotypes from their donor cells and mtDNA heteroplasmy could not be detected in any of the clones. The birth of healthy clones suggests that the haplotype difference between the cell and oocyte donor did not affect the pre- or post-implantation development of the bovine nuclear transfer derived embryos in our study. The results showed that well established nuclear transfer protocols could be useful in conserving endangered species. In conclusion, somatic cell banking can be suggested as a tool in conservation programmes of animal genetic resources.