Effect of culture methods on cumulus and oocyte morphology and meiotic competence of bovine oocytes from early antral follicles

Kątska-Książkiewicz, L.; Alm, H.

doi:10.5194/aab-48-562-2005

Cited by 3 publications

(3 citation statements)

References 25 publications

(49 reference statements)

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“…Cumulus expansion and maturation rates in the present study were comparable to the in studies utilizing EGF, FSH and LH as supplements during in vitro maturation of bovine oocytes 4,11 . This may show that our culture system may support oocyte maturation irrespective of hormonal supplementation because there are culture systems that maturation degree are similar to the level reached in our study 1,16 . When oocytes are removed from antral follicles, they begin spontaneous meiotic maturation, presumably due to release from inhibitory influences of substances in follicular fluid or produced by cumulus cells through gap junctions 13 .…”

Section: Discussionmentioning

confidence: 69%

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Sığır Oositlerinin Kumulus Ekspansiyonu ve Nükleer Maturasyon Üzerine Serum, Gonadotropinler, Epidermal Büyüme Faktörü ve Östradiol 17-Beta’nın Etkileri

Çevik¹,

Şen²,

Koçyiğit³

et al. 2009

Kafkas Univ Vet Fak Derg

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SummaryThe aim of the study was to evaluate the effects of follicle stimulating hormone (FSH), luteinizing hormone (LH), epidermal growth factor (EGF) and estradiol 17-β (E2) in maturation medium on nuclear maturation and cumulus expansion of cumulus-oocyte complexes (COCs) obtained from bovine ovaries. COCs were classified as good and poor quality grades based on cumulus investment. COCs were then subjected to in vitro maturation in TCM-199 in a humidified atmosphere of 5% CO2 in air at 38.5°C for a period of 24 h. The combination of the hormones added to the medium was as follows: T1=10% (v/v) heat inactivated Fetal Bovine Serum (FBS); T2=5 µg/ml bLH, 0.5 µg/ml bFSH and 10 ng/ml EGF; T3=FBS, bLH, bFSH and EGF; T4=1 µg/ml E2; T5=FBS and E2. Supplementation of maturation medium with the FSH, LH, EGF irrespective to supplementation with FBS stimulated expansion of cumulus around the oocytes compared to oocytes in other treatment groups (P<0.05). However there were no significant differences between treatment groups in terms of proportion of oocytes reached to metaphase II (M II) stage determined by nuclear staining. In general cumulus expansion degree increased with the quality of COCs in all treatment groups (P<0.05). In conclusion, hormone and growth factor supplementation (FSH, LH and EGF) stimulated cumulus cell expansion and maturation rate. Keywords: Bovine oocyte, IVM, EGF, Estradiol 17-β, Gonadotropins Sığır Oositlerinin Kumulus Ekspansiyonu ve Nükleer Maturasyon Üzerine Serum, Gonadotropinler, Epidermal Büyüme Faktörü ve Östradiol 17-Beta'nın Etkileri ÖzetÇalışmanın amacı, sığır ovaryumlarından elde edilen kumulus oosit komplekslerinin (KOK) nükleer olgunlaşma ve kumulus ekspansiyonu üzerine olgunlaşma medyumuna katılan folikül stimüle edici hormon (FSH), luteinleştirici hormon (LH), epidermal büyüme faktörü (EGF) ve östradiol 17-β (E2)'nin etkisini değerlendirmektir. KOK'lar oositlerin kumulus hücreleri ile kuşatılma derecesine göre iyi ve zayıf kaliteli olarak sınıflandırılmıştır. Sonrasında KOK'lar 24 saat süreyle TCM-199 medyumunda 38,5°C ve %5 CO2'li ortamda olgunlaşmaya bırakılmıştır. Medyuma eklenen hormonların kompozisyonu şu şekildedir; T1=10% (v/v) inaktif Fötal Sığır Serumu (FBS); T2=5 µg/ml bLH, 0.5 µg/ml bFSH ve 10 ng/ml EGF; T3=FBS, bLH, bFSH ve EGF; T4=1 µg/ml E2; T5=FBS ve E2. Olgunlaşma medyumuna FBS hariç FSH, LH ve EGF eklenmesinin diğer muamele grupları ile karşılaştırıldığında kumulus ekspansiyonunu artırdığı görülmüştür (P<0.05). Fakat nükleer boyama ile metafaz II (MII) safhasına ulaştığı belirlen oositlerin oranı muamele grupları arasında önemli bir fark göstermemiştir. Genel olarak tüm muamele gruplarında kumulus ekspansiyon derecesi KOK'ların kalitesiyle birlikte artış göstermiştir (P<0.05). Sonuç olarak hormon ve büyüme faktörü ilavesi kumulus ekspansiyonu ve olgunlaşma oranını stimüle etmiştir.

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Section: Discussionmentioning

confidence: 69%

“…Various approaches have been employed to improve the developmental competence of bovine oocytes in vitro 1 . Maturation media have been supplemented with various hormones, growth factors, sera, and follicular fluid in different reports for this aim [2][3][4][5][6][7] .…”

Section: Introductionmentioning

confidence: 99%

Sığır Oositlerinin Kumulus Ekspansiyonu ve Nükleer Maturasyon Üzerine Serum, Gonadotropinler, Epidermal Büyüme Faktörü ve Östradiol 17-Beta’nın Etkileri

Çevik¹,

Şen²,

Koçyiğit³

et al. 2009

Kafkas Univ Vet Fak Derg

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“…During maturation 12-14 h heat shock interrupted oocyte microtubule and microfilament texture (Roth and Hansen 2005) and spindle positioning (Ju et al 2005), diminished nuclear maturation (Roth and Hansen 2005;Rodrigues et al 2016), affection distribution of cortical granule (Maya-Soriano et al 2013) and reducing mitochondrial function in oocyte (Nabenishi et al 2012). Various approaches have been employed to improve maturation and developmental competence of cattle oocytes following in vitro fertilization (Katska-Ksiazkiewicz and Alm, 2005;Cevik et al, 2014). İn vitro maturation of cattle oocytes has been studied with different supplementation to culture media such as follicular fluid, co-culture (Moulavi et al, 2006), growth factors or gonadotropic hormones (Cevik et al, 2011).…”

Section: Bspublishersmentioning

confidence: 99%

Determination of 70 kDa Proteins for Different Temperatures in Cattle Oocytes Matured in vitro

Bener¹,

Kuran²

2019

Black Sea Journal of Engineering and Science

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This study was conducted to determine the effect of 36.5 (low) or 38.5 °C (conventional) culture temperatures during in vitro maturation (IVM) on 70 kDa proteins in cattle oocytes. In the study, cattle cumulus-oocyte complexes (COC) were subjected to IVM in bicarbonate-buffered TCM-199 supplemented with 10% FCS for 22 hours with a humidified 5% CO2 in air at either 36.5 or 38.5 °C culture temperatures. Maturation of COCs was determined according to fully cumulus expansion at the end of IVM. The cumulus cells of matured COCs were removed by vortexing in TCM-199 with HEPES buffered TCM-199 containing 0.1% hyaluronidase enzyme. Following, denuded oocytes were washed in 9.6 % phosphate-buffered saline (PBS) solution for protein isolation. Denuded oocytes (n = 100 for both groups) were centrifuged for 5 min at 300 g at 4 °C for PBS removed than the lysis buffer (100 μl) was added and shake 15 min on ice for 30 minutes. For protein isolation, the protein was centrifuged again at 13,000 g for 5 minutes and the protein samples were stored at-20 ° C. To increase the concentration of isolated protein samples, the samples were frozen at-80 °C for 24 hours and lyophilized at 0.140 hPa in a lyophilizer. Lyophilized protein samples were stored at-20 °C until SDS-PAGE analysis. The level of 70 kDa proteins in oocytes was determined by SDS-PAGE method. There were no significant differences between low (81.11%) or conventional (84.41%) incubation temperatures in IVM in terms of full cumulus expansion. The 70 kDa proteins band size in SDS-PAGE polyacrylamide gel image of cattle oocytes maturated at low culture temperature was weak compared to cattle oocytes maturated at conventional culture temperature. The results of this study may show that low incubation temperature during IVM decreases the amount of 70 kDa proteins in cattle oocytes.

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Animal reproduction biotechnology in Poland

Smorąg¹,

Kątska-Książkiewicz²,

Skrzyszowska³

et al. 2008

Int. J. Dev. Biol.

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The key research areas of the Department are: in vitro production of embryos, embryo cryopreservation, animal transgenesis, cloning, cytometric semen sexing and evaluation. Research has been focused on the in vitro production of animal embryos, including the development of complex methods for oocyte maturation, fertilization and embryo culture. Moreover, experiments on long-term culturing of late preantral and early antral bovine ovarian follicles have been developed. Studies on the cloning of genetically modified pigs with "humanized" immunological systems have been undertaken. A cloned goat was produced from oocytes reconstructed with adult dermal fibroblast cells. The novel technique of rabbit chimeric cloning for the production of transgenic animals was applied; additionally, the recipient-donor-cell relationship in the preimplantation developmental competences of feline nuclear transfer embryos has been studied. Regarding transgenic animal projects, gene constructs containing growth hormone genes connected to the mMt promoter were used. Modifications of milk composition gene constructs with tissue-specific promoters were performed. Moreover, pigs for xenotransplantation and animal models of human vascular diseases have been produced. Over the last 15 years, our flow cytometry research group has focused its work on new methods for sperm quality assessment and sex regulation. In the 1970s, our team initiated studies on embryo cryopreservation. As a result of vitrification experiments, the world's first rabbits and sheep produced via the transfer of vitrified embryos were born. KEY WORDS: animal cloning, transgenesis, sex regulation, cryopreservationThe Department of Biotechnology of Animal Reproduction in the National Research Institute of Animal Production was founded in 1950 as one of the institutes of the Ministry of Agriculture. During the first years of its history, the Department focused on practical issues related to insemination of cattle and horses. Prof. Wladyslaw Bielanski (Fig.1), the first Head of the Department, emphasized the practical side of research. In the mid 1960s, the Central Bank of Bull Semen was created for breeding evaluation, microbiological control of stored semen, and import and export of frozen semen.Prof. Stefan Wierzbowski, who became the Head of the Department in 1968, continued this practical approach and focused on the improvement of artificial insemination methods for various animal species, the development of semen preservation and evaluation methods, and animal behaviour and andrology.During the second half of the 1970s, the scope of the research in the Department was broadened to include aspects of female reproductive biology. Research on superovulation, embryo transfer and cryopreservation for several species of mammals was Int. J. Dev. Biol. 52: 151-155 (2008)

show abstract

Effect of culture methods on cumulus and oocyte morphology and meiotic competence of bovine oocytes from early antral follicles

Cited by 3 publications

References 25 publications

Sığır Oositlerinin Kumulus Ekspansiyonu ve Nükleer Maturasyon Üzerine Serum, Gonadotropinler, Epidermal Büyüme Faktörü ve Östradiol 17-Beta’nın Etkileri

Sığır Oositlerinin Kumulus Ekspansiyonu ve Nükleer Maturasyon Üzerine Serum, Gonadotropinler, Epidermal Büyüme Faktörü ve Östradiol 17-Beta’nın Etkileri

Determination of 70 kDa Proteins for Different Temperatures in Cattle Oocytes Matured in vitro

Animal reproduction biotechnology in Poland

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