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.

Distinct effect of fetal bovine serum versus follicular fluid on multipotentiality of human granulosa cells in in vitro condition

Biologicals ◽  
2018 ◽  
Vol 52 ◽  
pp. 44-48 ◽  
Author(s):  
Soudabe Yousefi ◽  
Jafar Soleimanirad ◽  
Kobra Hamdi ◽  
Laya Farzadi ◽  
Aalie Ghasemzadeh ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Follicular Fluid ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Distinct Effect ◽  
Human Granulosa Cells ◽  
Fetal Bovine

184 EFFECT OF CONCENTRATION AND EXPOSURE DURATION OF FETAL BOVINE SERUM ON PARTHENOGENETIC DEVELOPMENT OF PORCINE FOLLICULAR OOCYTES

2007 ◽  
Vol 19 (1) ◽  
pp. 208
Author(s):  
H. J. Kim ◽  
S. R. Cho ◽  
C. Y. Choe ◽  
S. H. Choi ◽  
D. S. Son ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Follicular Fluid ◽  
Bovine Serum ◽  
Exposure Duration ◽  
Cytochalasin B ◽  
Fetal Bovine Serum ◽  
Hatching Rate ◽  
Parthenogenetic Development ◽  
Maturation Medium ◽  
Fetal Bovine

The aim of the present experiment was to examine hatching rate as a testing tool of porcine embryo viability before early-stage embryo transfer, such as zygotes or 2-cell stage embryos. We evaluated the optimal concentrations and exposure durations of fetal bovine serum (FBS) on porcine parthenotes. Ovaries were obtained from prepubertal gilts at a local abattoir and brought to the laboratory in physiological saline with antibiotics at 30–33°C. The ovaries were washed and wiped, and then cumulus–oocytes complexes (COCs) in the follicular fluid were aspirated from surface-visible follicles (2–6 mm in diameter) with a 10-mL syringe fitted with an 18-gauge needle. After being washed 3 times with modified phosphate-buffered saline (DPBS; GIBCO, Grand Island, NY, USA) containing 0.3% BSA, the COCs were suspended in maturation medium, NCSU-23 containing 10% (v/v) porcine follicular fluid, 10 ng mL−1 epidermal growth factor (EFG; Sigma-Aldrich Corp., St Louis, MO, USA), 10 µg mL−1 follicular stimulating hormone (FSH; Sigma), 35 µg mL−1 luteinizing hormone (LH; Sigma), 1 mg mL−1 cysteine (Sigma, USA), 100 IU mL−1 penicillin G, and 100 µg mL−1 streptomycin sulfate (GIBCO). After 24 h, the COCs were transferred to the same medium without hormones. The oocytes matured for 48 h were denuded. The oocytes with a visible polar body were selected and returned to the maturation medium without hormones. After 65 h of maturation, oocytes were exposed to PBS with 7% ethanol (v/v) for 7 min, and then the oocytes were washed and treated in TCM-199 containing 5 µg mL−1 cytochalasin B (Sigma) for 5 h at 38.5°C in an atmosphere of 5% CO2 and 95% air with high humidity. After cytochalasin B treatment, the presumptive parthenotes were cultured in PZM-5 medium (IFP, Japan) and cleavage of the parthenotes was assessed at 72 h of activation. Normally cleaved parthenotes were cultured for 8 days to evaluate their ability to develop to the blastocyst and hatching stages. The FBS was added at Day 4 or 5 with concentrations of 2.5, 5, or 10%. The blastocyst rates ranged from 39.1 to 70% in each treatment. However, the hatching rate was dramatically decreased in the non-addition group. In this experiment, the developmental potential may be estimated before embryo transfer by an in vitro culture system up to the hatching stage. Table 1. Effect of concentration and exposure duration of FBS on parthenogenetic development of porcine follicular oocytes

scholarly journals Fetal bovine serum influences apoptosis and apoptosis-related gene expression in porcine parthenotes developing in vitro

Reproduction ◽  
2004 ◽  
Vol 127 (1) ◽  
pp. 125-130 ◽  
Author(s):  
Xiang-Shun Cui ◽  
Yu-Jeong Jeong ◽  
Hwa-Young Lee ◽  
Sun-Hong Cheon ◽  
Nam-Hyung Kim
Keyword(s):  
Gene Expression ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Blastocyst Stage ◽  
Related Gene ◽  
Embryo Viability ◽  
Cell Stage ◽  
Fetal Bovine ◽  
Apoptosis Related Gene

This study was conducted to determine the effects of polyvinyl alcohol (PVA), fetal bovine serum (FBS) and bovine serum albumin (BSA) on blastocoel formation, total cell number, apoptosis and Bcl-xL and Bak gene expression in porcine presumptive diploid parthenotes developing in vitro. The addition of 0.4% BSA to the culture medium enhanced the development of 2-cell or late 4-cell stage parthenotes to the blastocyst stage (P < 0.01) while FBS decreased the incidence of blastocoel formation. FBS also reduced the frequency of blastocysts developed from both 2-cell (P < 0.001) and late 4-cell (P < 0.05) embryos and increased the percentage of blastocysts undergoing apoptosis (P < 0.001). The relative abundance of Bcl-xL mRNA in presumptive diploid parthenotes in the control, PVA- and BSA-supplemented medium was similar to that of in vivo-derived embryos, but was significantly higher than in parthenotes cultured with FBS supplement (P < 0.05). Bak mRNA significantly increased at the blastocyst stage in FBS-supplemented cells (P < 0.01). These results suggest that apoptosis-related gene expression is significantly affected by FBS, and that this may result in alteration of apoptosis and embryo viability of porcine embryos developing in vitro.

Replacement of fetal bovine serum and FSH with buffalo follicular fluid in in vitro maturation of buffalo oocytes

1996 ◽  
Vol 45 (1) ◽  
pp. 245 ◽  
Author(s):  
S.K. Das ◽  
M.S. Chauhan ◽  
P. Palta ◽  
P.K. Katiyar ◽  
M.L. Madan
Keyword(s):  
Follicular Fluid ◽  
Bovine Serum ◽  
In Vitro Maturation ◽  
Fetal Bovine Serum ◽  
Fetal Bovine

In vitro maturation and fertilization, and subsequent development of buffalo (Bubalus bubalis) embryos: effects of oocyte quality and type of serum

1998 ◽  
Vol 10 (2) ◽  
pp. 173 ◽  
Author(s):  
M. S. Chauhan ◽  
S. K. Singla ◽  
P. Palta ◽  
R. S. Manik ◽  
M. L. Madan
Keyword(s):  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Cumulus Cells ◽  
Blastocyst Stage ◽  
Developmental Potential ◽  
Nuclear Maturation ◽  
Maturation Medium ◽  
Fetal Bovine ◽  
Grade 3

In Experiment 1, to determine the developmental potential of buffalo oocytes of different qualities, compact cumulus–oocyte complexes (COCs) with an unexpanded cumulus mass, and with homogeneous ooplasm were classified as Grade 1 (with 5 layers of cumulus cells) and Grade 2 less than 4 layers of cumulus cells). Grade-3 oocytes were either without cumulus cells or with expanded cumulus mass, and with irregular ooplasm. The oocytes were matured for 24 h at 38·5°C, 5% CO2 in air in maturation medium (10% fetal bovine serum (FBS) in TCM-199 supplemented with 5 µg mL-1 follicle stimulating hormone-P). The nuclear maturation and cleavage rates, and the proportion of cleaved embryos which developed to morula and blastocyst stage were in the order Grade 1>Grade 2>Grade 3 (P < 0·05). For Experiment 2, the maturation medium consisted of TCM-199 supplemented with one of the following sera at 10% concentration: (1) buffalo oestrus serum (BOS), (2) superovulated buffalo serum (SBS), (3) fetal bovine serum (FBS) and (4) steer serum (SS). After in vitro fertilization (IVF), the oocytes were co-cultured with buffalo oviductal epithelial cells in TCM-199 containing the respective sera at 10% concentration for the subsequent 9 days. The extent of cumulus expansion and nuclear maturation were not different among different groups. The cleavage rates were lower (P < 0·05) with FBS than with BOS, SBS and SS. The proportion of cleaved embryos which developed to blastocyst stage was higher (P < 0·05) with SBS than with BOS, FBS and SS.

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
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