scholarly journals EVALUASI OOSIT KAMBING HASIL IVM SEBAGAI SALAH SATU FAKTOR PENENTU KEBERHASILAN DALAM AKTIVASI PARTENOGENESIS

el–Hayah ◽  
2012 ◽  
Vol 2 (1) ◽  
Author(s):  
Kholifah Holil
Keyword(s):  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Polar Body ◽  
Follicle Stimulating Hormone ◽  
Fetal Bovine ◽  
Stimulating Hormone

<p>Aktivasi partenogenesis merupakan salah satu tehnik aktivasi oosit untuk menghasilkan embrio tanpa kontribusi dari sperma. Salah satu faktor yang menentukan keberhasilan tehnik ini adalah pada ketersediaan oosit yang berkualitas. Oleh karena itu penelitian ini bertujuan untuk mengevaluasi oosit kambing hasil IVM yang dapat digunakan untuk kepentingan dalam aktivasi partenogenesis tersebut.<br />Sampel yang digunakan dalam penelitian ini adalah oosit yang diaspirasi dari folikel ovarium kambing yang diambil dari RPH Sukun Malang. Oosit di IVM selama 24 jam dan selama 27 jam dalam medium TCM-199 yang ditambah dengan fetal bovine serum (FBS), follicle-stimulating hormone (FSH) dan lutheinizing hormone (LH) dan diinkubasi pada suhu 38,5oC, 5% CO2. Pada jam ke 24 dan jam ke 27 setelah IVM dilakukan pengamatan yang meliputi ekspansi sel-sel kumulus dan keberadaan polar body I (PB-I) pada jam ke 30. <br />Berdasarkan hasil penelitian menunjukkan bahwa oosit kambing  yang di IVM  sampai jam ke 24 belum menunjukkan adanya ekspansi sel-sel kumulus kualitas 2 dan penampakan PB-I. Hasil tersebut berbeda dengan oosit yang di IVM sampai jam ke 27. Pada pengamatan jam ke 27 ini menunjukkan bahwa terdapat 77,87% oosit yang sel-sel kumulusnya berekspansi (kualitas 2) dan PB-I yang nampak sebesar 95,32%.  Dengan demikian maka dapat disimpulkan bahwa oosit kambing hasil IVM  baru dapat digunakan untuk keperluan lebih lanjut khususnya untuk keperluan aktivasi partenogenesis pada jam ke 27.</p><p>Kata kunci: evaluasi, aktivasi partenogenesis, oosit kambing hasil IVM <br /><br /></p>

scholarly journals Fetal bovine serum is associated with polar body degeneration after in vitro maturation of bovine oocytes

2018 ◽  
Vol 68 (3) ◽  
pp. 279
Author(s):  
B. MACÍAS-GARCÍA ◽  
S. MACEDO ◽  
A. ROCHA ◽  
L. GONZÁLEZ-FERNÁNDEZ
Keyword(s):  
Bovine Serum ◽  
Culture Medium ◽  
Fetal Bovine Serum ◽  
Polar Body ◽  
Chromatin Conformation ◽  
Prolonged Incubation ◽  
Vitro Fertilization ◽  
Fetal Bovine ◽  
Bovine Oocytes

In vitro fertilization (IVF) in cattle is commonly used worldwide. Although extensive research has been conducted using different additives in the different IVF steps, little is known regarding how protein type may affect bovine oocytes during the fertilization period. In addition, unlike Tissue Culture Medium 199 (TCM), fertilization medium may induce oocytes’ chromatin degeneration during prolonged incubation in the horse (Modified Whitten’s medium). Thus, in the present work TCM-199 supplemented with either 7 mg/ml of Bovine Serum Albumin (TCM+BSA) or 10% Fetal Bovine Serum (v/v; TCM+FBS) was used. Bovine oocytes were matured in vitro and placed in the previously mentioned media for further 18 hours, in the absence of added sperm (sham fertilization) and their chromatin conformation was evaluated. After IVM, 78.9% of the initial oocytes had reached the MII stage. After sham fertilization, 58.6% of the oocytes in TCM+BSA while just 28.3% in TCM+FBS maintained the MII chromatin conformation (p < 0.05). Subsequent experiments run using PB extruded oocytes and incubated in TCM+BSA and TCM+FBS during sham fertilization, demonstrated that FBS was consistently associated with polar body dissolution or degeneration.

131 In vitro nuclear maturation and blastocyst developmental rates after intracytoplasmic sperm injection of equine oocytes held for 24 h at room temperature in Tyrode’s albumin lactate pyruvate-Hepes (TALP-h) or in a commercial embryo holding medium

2021 ◽  
Vol 33 (2) ◽  
pp. 173
Author(s):  
A. F. Bragulat ◽  
A. Gambini ◽  
M. B. Rodriguez ◽  
O. Briski ◽  
C. Alonso ◽  
...  
Keyword(s):  
Embryo Development ◽  
Intracytoplasmic Sperm Injection ◽  
Bovine Serum ◽  
Room Temperature ◽  
Fetal Bovine Serum ◽  
Polar Body ◽  
Immature Oocytes ◽  
Nuclear Maturation ◽  
First Polar Body ◽  
Fetal Bovine

The interest in equine intracytoplasmic sperm injection (ICSI) for commercial and research applications has rapidly increased. Shipping immature oocytes at room temperature has been proven successful, and to identify the optimal conditions for holding oocytes, several mediums are being tested. The aim of this study was to compare the effect of holding equine oocytes in Tyrode’s albumin lactate pyruvate-Hepes (TALP-h, Bavister and Yanagimachi 1977 Biol. Reprod. 16, 228-237) medium or in commercial embryo holding medium (EHM, Syngro® Holding) on invitro nuclear maturation rates and pre-implantation embryo development after ICSI. Cumulus–oocyte complexes (COCs) were recovered from ovaries of slaughtered mares and assigned randomly in 2-mL cryovials with TALP-h or EHM, with a maximum of 30 oocytes per cryovial. COCs were shipped to the ICSI laboratory at 20 to 25°C for 24 to 28h followed by IVM for 24h in a humidified atmosphere of 5% CO2 in air at 38.5°C. Maturation medium was TCM-199 with 10% fetal bovine serum, 1μL mL−1 insulin-transferrin-selenium, 1mM sodium pyruvate, 100mM cysteamine, and 0.1mg mL−1 FSH. After mechanical cumulus cell removal, nuclear maturation rate was assessed using a stereomicroscope. Oocytes with an intact oolemma and extrusion of the first polar body (PB) were classified as mature, oocytes without a visible PB were considered immature, and oocytes without an intact oolemma were considered degenerate. Matured oocytes were subjected to ICSI without piezo-drill system (one proved stallion) in 20-μL droplets of TALP-h with a 7-μm glass sharp micropipette in an inverted microscope (Nikon Eclipse TE-300 microscope) using hydraulic micromanipulators (Narishige, Medical Systems). Presumptive ICSI zygotes were cultured in DMEM F12/Global Total® with 6% fetal bovine serum for 9 days at 38.5°C in a humidified atmosphere of 5% O2 and 5% CO2 in air. On Day 5 of culture, cleavage was recorded and medium was refreshed. Blastocysts rates were recorded on Day 7 and 9 of culture. Invitro nuclear maturation rates are shown in Table 1. We observed a significantly higher proportion of immature oocytes in the EHM group compared with the TALP-h group. After ICSI of some matured oocytes of each group, no significant differences were observed in cleavage or blastocyst rate (Table 1). Our results suggest that either TALP-h or commercial embryo holding medium are suitable for oocyte shipping and to support blastocyst development after ICSI. Table 1. Invitro nuclear maturation rates and pre-implantation embryo development after intracytoplasmic sperm injection (ICSI) Maturation rates Medium Oocytes Mature [n (%)] Immature [n (%)] Degenerate [n (%)] TALP-h 315 173 (54.9) 26 (8.3)a 116 (36.8) EHM 273 132 (48.4) 55 (20.1)b 86 (31.5) Total 588 305 (51.9) 81 (13.8) 202 (34.4) Embryo development ICSI (n) Cleaved [n (%)] Day 7 Blastocyst [n (%)] Day 9 Blastocyst [n (%)] TALP-h 35 23 (65.7) 7 (20) 9 (25.7) EHM 26 19 (73.1) 3 (11.5) 5 (19.2) Total 61 42 (68.9) 10 (16.4) 14 (23) a,bDifferent superscript letters indicate statistical significance (Fisher’s exact test, P&lt;0.05).

41 THE EFFECT OF ULTRARAPID VITRIFICATION FOR SHEEP OOCYTE VIABILITY

2017 ◽  
Vol 29 (1) ◽  
pp. 128
Author(s):  
M. Toishibekov ◽  
Y. Toishibekov ◽  
M. Yermekova
Keyword(s):  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Polar Body ◽  
Control Group ◽  
Polar Bodies ◽  
Chi Squared ◽  
Fetal Bovine ◽  
Second Polar Body ◽  
Cryopreservation Techniques

Advances in reproduction technologies, such as in vitro maturation, IVF, and in vitro culture, have stimulated research for efficient cryopreservation techniques for mammalian oocytes. It is well known that the oocyte is the largest cell of an animal’s body and as such, is full of water and, in many species, fat, making it difficult to cryopreserve. The objective of this work was to study the effect of vitrification for cryopreservation of the metaphase II plate (MPII) of sheep oocytes. In our experiment, we used the Vit-Master™ (MTG, Bruckberg, Germany). Ovaries from 19 ewes of Kazakh Arkharo-Merino breed were acquired after slaughter and maintained at 37°C in TCM-199. The maturation medium was TCM-199, containing 1 mM of glutamine, 10% fetal bovine serum, 5 μg mL−1 FSH, 5 μg mL−1 LH, 1 μg mL−1 oestradiol, 0.3 mM sodium pyruvate, and 100 mM cysteamine. The oocytes were incubated in 400 μL of medium in 4-well dishes covered with mineral oil. The IVM conditions were 5% CO2 in humidified air at 39°C for 24 h. Then, oocytes were placed for 10 min in medium with Hoechst 33342 (3 μg mL−1) and cytochalasin B (7 μg mL−1) to facilitate enucleation of the MPII with a minimum volume of ooplasm. The MPII plates were divided into 2 groups: the vitrification group was exposed to vitrification media containing 1.12 M ethylene glycol (ET) + 0.87 M ME2SO for 5 min and was exposed in vitrification media containing 2.24 M ET + 1.75 M ME2SO for 5 min, and then in vitrification solution containing 4.48 M ET + 40% ME2SO + 0.25 M sucrose for 30 s. Oocytes were loaded into a cryoloop and using negative pressure of liquid nitrogen in the chamber for freezing with the VIT-Master. Oocytes were thawed in a 25°C water bath and then placed in TCM-199 at 20% fetal bovine serum. After 15 min of incubation, the oocytes were activated for extrusion of the second polar body in 1 mg mL−1 Ca ionophore for 5 min and washed for 5 min followed by 4 h in 6-DMAP (0.12 mM) + cycloheximide (0.6 μg mL−1). After activation, the MPII were washed and cultured for 20 h. The control group received the same treatment but were not vitrified. Differences between the experimental groups were tested using Chi-squared test. Our research showed that expulsion of the second polar body after activation was observed in more than 59.7% of the MPII that were not vitrified (control group), whereas 37.7% of vitrified plates had expulsion of polar bodies (P < 0.05). These preliminary studies showed that it is possible to vitrify MPII plates. On the other hand, the drastic reduction of the volume of the sheep oocytes might make cryopreservation possible with greater efficiency.

Treatment of fetal bovine serum improves early development of porcine embryos by alleviating oxidative stress

2014 ◽  
Author(s):  
Seon-A Choi ◽  
Seong-Eun Mun ◽  
Pil-Soo Jeong ◽  
Hae-Jun Yang ◽  
Seung-Bin Yoon ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Early Development ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Fetal Bovine

МОДЕЛИРОВАНИЕ СИСТЕМ ЭКСТРАКОРПОРАЛЬНОГО СОЗРЕВАНИЯ ООЦИТОВ КРУПНОГО РОГАТОГО СКОТА С ИСПОЛЬЗОВАНИЕМ СТРУКТУРНЫХ КОМПОНЕНТОВ ОВАРИАЛЬНЫХ ФОЛЛИКУЛОВ *

2019 ◽  
pp. 20-22
Author(s):  
T.I. KUZMINA ◽  
I.V. CHISTYAKOVA
Keyword(s):  
Granulosa Cells ◽  
Follicular Fluid ◽  
Bovine Serum ◽  
Egg Quality ◽  
Fetal Bovine Serum ◽  
Reproductive Technologies ◽  
Blastocyst Stage ◽  
Preimplantation Embryos ◽  
Cultural Systems ◽  
Fetal Bovine

Создание эффективной унифицированной системы дозревания донорских ооцитов обеспечит повышение результативности инновационных клеточных репродуктивных технологий. В исследовании проведен сравнительный мониторинг показателеймейотического созревания ооцитов коров, созревших в различных системах, дополненных структурными компонентами фолликулов (СКФ стенки фолликулов, клетки гранулезы, белки) и фолликулярной жидкостью,а также потенций к развитию из них доимплантационных эмбрионов. Анализу подверглись ооциты, прокультивированные в следующих системах:среда ТС199 с добавлением 10 фетальной бычьей сыворотки (ФБС), 50 мкг/мл эстрадиола, 10 мкг/мл лютеинизирующего гормона (ЛГ), 10 мкг/мл фолликулостимулирующего гормона (ФСГ) среда ТС199 с 10 эстральной сывороткой коров среда ТС199 с 50 жидкости из фолликулов диаметром 9 мм среда ТС199 с добавлением белков фолликулярной жидкости молекулярной массой 65 кДасреда ТС199 с 10 ФБС и 1106 клеток гранулезы среда ТС199 с 10 ФБС и тканью фолликула. В культуральные среды ко всем исследованным группам ооцитов добавляли антибиотики. Использование CКФ обеспечило значительное снижение доли ооцитов с дегенерированным хроматином, что способствовало увеличению уровня доимпланационных эмбрионов на стадии бластоцисты. Так, доля бластоцист, развившихся из ооцитов, созревших в среде со стенками фолликулов,составила43,5. В этой же группе выявлен минимальный уровень дегенерированных зародышей (6,45). Полученные данные предлагается использовать при моделировании систем дозревания ооцитов коров с целью повышения качества яйцеклеток.The creation of an effective unified maturation system of donor oocytes provides an increase in the efficiency of innovative cellular reproductive technologies. The comparative analysis of the meiotic maturation indicators of bovine oocytes, which were matured in different cultural systems modified by follicular structural components (FSC follicular walls, granulosa cells, proteins) and follicular fluid, as well as the potential for preimplantation embryonic development were evaluated in this study. Oocytes matured in following cultural systems: medium TC199 supplemented with 10 fetal bovine serum and 50 g/ml of estradiol, 10 g/ml of luteinizing hormone (LH), 10 g/ml of folliclestimulating hormone (FSH) medium TC199 with 10 estrous cow serum medium TC199 with 50 liquid from follicles with a diameter of 9 mm medium TC199 supplemented with the follicular fluid proteins with molecular weight 65 kDa medium TC199 with 10 fetal bovine serum and 1106 granulosa cells medium TC199 with the addition of 10 fetal bovine serum and follicle tissues were analyzed. Antibiotics were added to cultural media of all experimental groups of oocytes. The usage of FSC ensured the decrease in the proportion of oocytes with degenerated chromatin, which contribute the rise of the level of preimplantation embryos at the blastocyst stage. Thus, the proportion of blastocysts developed from oocytes matured in medium supplemented with follicular walls was 43.5. In the same experimental group, the number of degenerated embryos was 6.45. The obtained data are supposed to be used for modeling the cultural systems of cow oocytes in order to improve the egg quality.

scholarly journals Effect of fetal bovine serum on erythropoietin receptor expression and viability of breast cancer cells

2020 ◽  
Vol 27 (2) ◽  
pp. 653-658
Author(s):  
Guan-Young Teo ◽  
Abdullah Rasedee ◽  
Nagi. A. AL-Haj ◽  
Chaw Yee Beh ◽  
Chee Wun How ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Bovine Serum ◽  
Breast Cancer Cells ◽  
Fetal Bovine Serum ◽  
Erythropoietin Receptor ◽  
Receptor Expression ◽  
Fetal Bovine

scholarly journals Prevalence of Bovine Viral Diarrhea Virus Genotypes and Antibody against those Viral Genotypes in Fetal Bovine Serum

1998 ◽  
Vol 10 (2) ◽  
pp. 135-139 ◽  
Author(s):  
Steven R. Bolin ◽  
Julia F. Ridpath
Keyword(s):  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Neutralizing Antibody ◽  
Bovine Viral Diarrhea ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Fetal Bovine ◽  
Viral Diarrhea Virus

One thousand lots of pooled fetal bovine serum (FBS) were tested for contamination with bovine viral diarrhea virus (BVDV) and/or for contamination with neutralizing antibody against BVDV. Noncytopathic or cytopathic BVDV was isolated from 203 lots of FBS. Analysis of the viral isolates identified 115 type 1 and 65 type 2 BVDV isolates. An additional 23 virus isolates were mixtures of >2 BVDV isolates and were not classified to viral genotype. Further characterization of the type 1 viruses identified 51 subgenotype 1a and 64 subgenotype 1b BVDV isolates. Viral neutralizing antibody was detected in 113 lots of FBS. Differential viral neutralization indicated that type 1 BVDV induced the antibody detected in 48 lots of FBS and type 2 BVDV induced the antibody detected in 16 lots of FBS.

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