Effects of two different extenders on the storage of cock semen at +5 degrees C

Total 12 cocks kept in individual cages were used in this study. Two times a week, total 24 ejaculates were collected from each cock and semen was pealed for spermatological evaluations. Spermatological tests were semen volume, mass activity, spermatozoa motility, spermatozoa concentration, live-dead spermatozoa rate and abnormal spermatozoa rate. The values of these characters were 3.29 +/- 1.19 ml, 3.29 +/- 0.62 (+), 85.83 +/- 6.19 %, 4.34 +/- 1.69 x 10(9)/ml, 83.34 +/- 6.43 %, 11.83 +/- 0.96 % respectively

The effects of various BSA levels in different media on development in in vitro culture of mouse embryos

Various BSA (bovine serum albumin) levels and media effects on development of 2-cell stage mouse embryos in in vitro culture were investigated in this 2 stage study. In the first stage, 1400 2-celled mouse embryos were cultured in 2 various BSA-bearing culture media for 96 h and directly observed microscopically. In the second stage, 1024 2-celled mouse embryos cultured for the same period and cell numbers (nuclei of cells) were determined by staining techniques

Effects of maturation and fertilization times on pronuclear development of bovine oocytes fertilized in vitro

Primer oocytes (n=520) collected from ovaries of slaughtered cows were divided into 3 groups, and matured at 39 degrees C with 95-100% humidity and under a gas mixture of 5% O-2, 5% CO2 and 90% N-2 for 22, 24 and 26 hours. Modified Parker's Medium (MPM) supplemented with FSH and 20% ECS was used as maturation medium. At the end of these periods, oocytes (22 h.=122, 24 h.=143 and 26 h.=255) were taken into Tyrode-lactate medium supplemented with BSA, heparin and PHE, and frozen-thawed bull semen was add after swim-up procedure (10 mu congruent to 400.000 spermatozoa/30-35 oocytes). Oocytes matured in 3 groups were kept with spermatozoa for 14, 16, 18, 20, 22 and 24 hours. At the end of these time periods all oocytes were fixed, stained and examined by phase-contrast microscope

Development of in vitro derived sheep embryos to the blastocyst stage

The objective of the present study was to mature and fertilize primary sheep oocytes in vitro and to develop them to the blastocyst stage Ovaries from slaughtered Kivircik ewes were used The ovaries were transported to the laboratory in a vacuum flask in which 30 35 C PBS (phosphate buffered saline) was included The cumulus oocyte complexes were collected by rupturing and washing the follicle wall with oocyte washing medium They were then selected under a stereo microscope and maturated for 23 24 hours in medium 199 supplemented with sodium pyruvate follicle stimulating hormone (FSH) luteinizing hormone (LH) and 10 / fetal calf serum (FCS) under 5 / CO2 atmosphere and at 38 5 C temperature Semen collected and pooled from three Kivircik rams by electro ejaculation and prepared for in vitro fertilization by Percoll gradient method was incubated for 20 21 hours with oocytes in BSOF medium supplemented with 2% sheep estrous serum (SES) (0 8x10(6) spermatozoon/ml) Presumptive zygotes were cultured in SOF medium for 8 days in synthetic oviduct medium (SOF) under an atmosphere of 5% CO2 5% O-2 and 90% N-2 Glucose (1 5mM) was added to the culture medium on Day

Effects of equilibration media and co-culture on vitrification of sheep blastocysts derived in vitro

İki vitrifikasyon protokolünün, koyun ovidukt epitel hücreleriyle ko-kültürlü (CC) ve ko- kültürsüz (C) ortamda in vitro üretilmiş olan koyun blastosistlerinin yaşayabilirliği üzerindeki etkileri araştırıldı. Oositler, TCM 199 medyumunda 24 saat süreyle olgunlaştırıldı, 20 saat fertilize edildi ve 9 gün süreyle sentetik ovidukt sıvısı (SOF) medyumunda kültüre edildi. Embriyolar ilk olarak, %20 etilen glikol (EG) veya % 10 gliserol (G) den oluşan iki farklı medyumda ekilibrasyona bırakıldılar. İkinci ekilibrasyon amacıyla % 20 etilen glikol + % 10 gliserol içinde ve 5 dakika süreyle tutuldular. Ardından, % 25 etilen glikol + % 25 gliserol’den oluşan vitrifikasyon solüsyonu içerisinde 30 saniye tutulan embriyolar, derhal sıvı azot içerisine batırılarak donduruldu. Çözdürme işleminin ardından embriyolar, 0.25 M sukroz içine 5 dakika bekletildi, Hepes tamponlu sentetik ovidukt sıvısı (HSOF) içinde yıkandı ve 24 saat süreyle SOF içerisinde in vitro kültüre bırakıldı. Yarıklanma oranları C grubunda %75.2, CC grubunda %74.2, blastosist oranları C'de %14.4, CC grubunda ise %17.1 oldu. Dondurulmuş-çözdürülmüş blastosistlerin in vitro kültüründen sonra canlılık oranları C-EG, CC-EG, C-G ve CC-G gruplarında sırasıyla; %62.1, %38.4, %30.2 ve %39.3 oldu. Bu çalışmada, koyun embriyolarının vitrifikasyonunda; ilk ekilibrasyon medyumu olarak gliserol yerine etilen glikol kullanılmasının faydalı olduğu buna karşılık, koyun ovidukt epiteli hücreleri ile ko-kültürün etkili olmadığı saptandı

pH sensitive functionalized hyperbranched polyester based nanoparticulate system for the receptor-mediated targeted cancer therapy

The aim of this study is to develop a novel folic acid conjugated, DL-lysine modified, PEGylated, the 3rd generation hyperbranched polymer (HBP-PEG-Lys-FA) for use in receptor-mediated therapy. 5-fluorouracil, model anti-cancer drug, loaded nanoparticles were found an average size of 177 nm with loading efficiency of 23.18%. In vitro drug release studies demonstrated that nanoparticles showed pH-dependent release. HBP-PEG-Lys-FA were efficiently taken up by HeLa cells and specificity of targeted nanoparticles to folate receptors of cells was proved. It was concluded that the HBP-PEG-Lys-FA nanoparticles can provide an advantage on delivering of the drug efficiently into the cytosol for cancer therapy

Reproductive performance of first cloned Anatolian Grey Cattle produced by frozen cells from National Animal Gene Bank

European Biotechnology Congress -- MAY 15-18, 2014 -- Lecce, ITALY[No Abstract Available

Estimation of the potential fertility based upon non-return rates of bulls: Using polyacrylamide gel instead of cervical mucus in the sperm penetration test

In the present study, we aimed to develop a polyacrylamide gel that could be used instead of bovine cervical mucus in the cervical mucus penetration test (CMPT) to obtain coherent and replicable results in bulls. The frozen semen samples of six Holstein bulls, which were divided into two fertility groups as low and high according to their non-return rate (NRR), were used. In this study, the modified CMPT (mCMPT) was carried out within 0.25 mL transparent plastic straws with an inner diameter 1.7 mm. The penetration ability of spermatozoa to bovine cervical mucus and to polyacrylamide gels swollen with two different solutions [NaCl (G1) and PBS (G2)] was compared. For the penetration test, the straws filled with cervical mucus and both gels were dipped into thawed semen samples and incubated at 37 T for 15 min. After the incubation, straws were frozen in liquid nitrogen vapour and stored at -20 degrees C. On the evaluation day, the frozen straws were cut at 1.5-1.75 cm (penetration distance range = PDRI), 3.25-3.5 cm (PDR2) and 5.0-5.25 cm (PDR3), beginning from open-end of the straws. The separated frozen parts were then immediately transferred onto special counting slides by pushing with a mandrel and left to thaw. Thawed samples were covered with cover glass and penetrated spermatozoa in these parts were counted. The relation between the results and fertility of bulls was determined

Effects of serum starvation and ionomycin activation on the development of somatic cell nuclear transfer embryos in sheep

Donör hücrelerin senkronizasyonu ve yeniden yapılandırılmış oositlerin aktivasyonu, somatik hücre klonlamasındaki başarı oranını etkileyen önemli faktörlerdir. Bu çalışmada, Kıvırcık koyunlarında somatik hücre nükleer transferinde serum açlığının donör hücre senkronizasyonuna ve ionomisinin yeniden yapılandırılmış oositlerin aktivasyonuna olan etkileri araştırılmıştır. Donör hücre olarak kullanılan kumulus hücreleri mezbahadan alınan koyun ovaryumundan elde edildi. Bu hücreler çalışmada 4 gün serum açlığına bırakılarak (%0.5 FCS; SS) veya serum açlığına bırakılmadan (%10 FCS; S) kullanıldı. Yeniden yapılandırma sonrası oositlerin aktive olması için ionomisinle 5 dk ve 6-dietilaminopurin ile 3 saat (I+) veya sadece 6-dietilaminopurin ile 3 saat (I-) inkübe edildi. İn vitro kültürün ikinci gününde, bölünmüş embriyolar (n=44) senkronize edilmiş alıcı koyunlara (n=10) transfer edildi. Embriyoların yarıklanma oranları SS/I+, S/I+, SS/I- ve S/I- gruplarında sırasıyla % 37.3; % 44.1; % 34.6 ve % 44.7 bulundu. 18. günde yapılan progesteron analizine göre kan progesteron seviyesi >1 ng/ml olan taşıyıcı koyun oranları sırasıyla % 33.3; %50.0; %50.0 ve %100.0 olarak tespit edildi. Gruplardan da sadece S/I- grubundan tek bir gebelik 40. günden sonra gelişimine devam edebildi. Ancak bu klon kuzu (transfer edilen embriyo sayısına göre, %7.1) doğuma 10 gün kala maternal bir problem (torsiyo uteri) sebebiyle öldü. Bu çalışmanın sonucunda koyunlarda somatik hücre nükleer transferinde serum açlığı ile somatik hücre senkronizasyon ve ionomisin ile oosit aktivasyon yöntemlerine gerek olmadan klon yavru elde edilebileceği ortaya konulmuştur