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.

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.

scholarly journals Supplementation of fetal bovine serum alters histone modification H3R26me2 during preimplantation development of in vitro produced bovine embryos

2015 ◽  
Vol 35 (7) ◽  
pp. 605-612
Author(s):  
Daniel R. Arnold ◽  
Carolina A.P. Corrêa ◽  
Laura L.G. Lorena ◽  
Roberta C. Gaspar ◽  
Guilherme F. Rossi ◽  
...  
Keyword(s):  
Histone Modification ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Bovine Embryos ◽  
In Vitro Production ◽  
Cell Stage ◽  
Morula Stage ◽  
Fetal Bovine

Abstract In vitro production (IVP) of bovine embryos is not only of great economic importance to the cattle industry, but is also an important model for studying embryo development. The aim of this study was to evaluate the histone modification, H3R26me2 during pre-implantation development of IVP bovine embryos cultured with or without serum supplementation and how these in vitro treatments compared to in vivo embryos at the morula stage. After in vitro maturation and fertilization, bovine embryos were cultured with either 0 or 2.5% fetal bovine serum (FBS). Development was evaluated and embryos were collected and fixed at different stages during development (2-, 4-, 8-, 16-cell, morula and blastocyst). Fixed embryos were then used for immunofluorescence utilizing an antibody for H3R26me2. Images of stained embryos were analyzed as a percentage of total DNA. Embryos cultured with 2.5% FBS developed to blastocysts at a greater rate than 0%FBS groups (34.85±5.43% vs. 23.38±2.93%; P<0.05). Levels of H3R26me2 changed for both groups over development. In the 0%FBS group, the greatest amount of H3R26me2 staining was at the 4-cell (P<0.05), 16-cell (P<0.05) and morula (P<0.05) stages. In the 2.5%FBS group, only 4-cell stage embryos were significantly higher than all other stages (P<0.01). Morula stage in vivo embryos had similar levels as the 0%FBS group, and both were significantly higher than the 2.5%FBS group. These results suggest that the histone modification H3R26me2 is regulated during development of pre-implantation bovine embryos, and that culture conditions greatly alter this regulation.

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.

Successful piglet production by IVF of oocytes matured in vitro using NCSU-37 supplemented with fetal bovine serum

2006 ◽  
Vol 65 (2) ◽  
pp. 374-386 ◽  
Author(s):  
Misae Suzuki ◽  
Koji Misumi ◽  
Manabu Ozawa ◽  
Junko Noguchi ◽  
Hiroyuki Kaneko ◽  
...  
Keyword(s):  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Fetal Bovine

scholarly journals Stimulatory Role of Glycated Fetal Bovine Serum Along with Iron on In Vitro Production of Insulin by Differentiated Mouse Embryonic Stem Cells

2011 ◽  
Vol 57 (4) ◽  
pp. 356-361
Author(s):  
Ikuo Nishigaki ◽  
Gowri Rangasamy Gunassekaran ◽  
Panjan Nagappan Venkatesan ◽  
Mandupal Chaco Sabu ◽  
Sabu Priya ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Embryonic Stem ◽  
In Vitro Production ◽  
Fetal Bovine ◽  
Vitro Production

scholarly journals Fetal Bovine Serum-Derived Extracellular Vesicles Persist within Vesicle-Depleted Culture Media

2018 ◽  
Vol 19 (11) ◽  
pp. 3538 ◽  
Author(s):  
Brandon Lehrich ◽  
Yaxuan Liang ◽  
Pooya Khosravi ◽  
Howard Federoff ◽  
Massimo Fiandaca
Keyword(s):  
Cell Growth ◽  
Extracellular Vesicles ◽  
Bovine Serum ◽  
Culture Media ◽  
Fetal Bovine Serum ◽  
Cellular Growth ◽  
Primary Astrocyte ◽  
Fetal Bovine ◽  
Cell Growth And Viability

It is known that culture media (CM) promotes cellular growth, adhesion, and protects explanted primary brain cells from in vitro stresses. The fetal bovine serum (FBS) supplement used in most CM, however, contains significant quantities of extracellular vesicles (EVs) that confound quantitative and qualitative analyses from the EVs produced by the cultured cells. We quantitatively tested the ability of common FBS EV-depletion protocols to remove exogenous EVs from FBS-supplemented CM and evaluated the influence such methods have on primary astrocyte culture growth and viability. We assessed two methodologies utilized for FBS EV removal prior to adding to CM: (1) an 18-h ultracentrifugation (UC); and (2) a commercial EV-depleted FBS (Exo-FBS™). Our analysis demonstrated that Exo-FBS™ CM provided the largest depletion (75%) of total FBS EVs, while still providing 6.92 × 109 ± 1.39 × 108 EVs/mL. In addition, both UC and Exo-FBS™ CM resulted in poor primary astrocyte cell growth and viability in culture. The two common FBS EV-depletion methods investigated, therefore, not only contaminate in vitro primary cell-derived EV analyses, but also provide a suboptimal environment for primary astrocyte cell growth and viability. It appears likely that future CM optimization, using a serum-free alternative, might be required to advance analyses of cell-specific EVs isolated in vitro.

scholarly journals Profiling of human neural crest chemoattractant activity as replacement of fetal bovine serum for in vitro chemotaxis assays

2021 ◽  
Author(s):  
Xenia Dolde ◽  
Christiaan Karreman ◽  
Marianne Wiechers ◽  
Stefan Schildknecht ◽  
Marcel Leist
Keyword(s):  
Neural Crest ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Developmental Toxicity ◽  
Video Tracking ◽  
Large Panel ◽  
Human Proteins ◽  
Fetal Bovine ◽  
Targeted Approach

Fetal bovine serum (FBS) is the only known stimulus for migration of human neural crest cells (NCCs). Non-animal chemoattractants are desirable for the optimization of chemotaxis assays to be incorporated in a test battery for reproductive and developmental toxicity. We confirmed here in an optimized transwell assay that FBS triggers directed migration along a concentration gradient. The responsible factor was found to be a protein in the 30-100 kDa size range. In a targeted approach, we tested a large panel of serum constituents known to be chemotactic for NCCs in animal models (e.g. VEGF, PDGF, FGF, SDF-1/CXCL12, ephrins, endothelin, Wnt, BMPs). None of the corresponding human proteins showed any effect in our chemotaxis assays based on human NCCs. We then examined in a broad screening approach, whether human cells would produce any factor able to trigger NCC migration. We found that HepG2 hepatoma cells produced chemotaxis-triggering activity (CTA). Using chromatographic methods and by employing the NCC chemotaxis test as bioassay, the responsible protein was enriched by up to 5000-fold. We also explored human serum and platelets as direct source, independent of any cell culture manipulations. A CTA was enriched from platelet lysates several thousand-fold. Its temperature and protease-sensitivity suggested a protein component. The capacity of this factor to trigger chemotaxis was confirmed by single-cell video-tracking analysis of migrating NCCs. The human CTA characterized here may be employed in the future for the setup of assays testing for the disturbance of directed NCC migration by toxicants.

scholarly journals Influence of recombinant hematopoietins and of fetal bovine serum on the globin synthetic pattern of human BFUe

Blood ◽  
1990 ◽  
Vol 76 (6) ◽  
pp. 1150-1157 ◽  
Author(s):  
AR Migliaccio ◽  
G Migliaccio ◽  
M Brice ◽  
P Constantoulakis ◽  
G Stamatoyannopoulos ◽  
...  
Keyword(s):  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Mrna Level ◽  
Fetal Hemoglobin ◽  
Mrna Levels ◽  
Gm Csf ◽  
Gamma Globin ◽  
Fetal Bovine ◽  
Globin Synthesis

Abstract We have studied the effects of recombinant hematopoietic growth factors, granulocyte-macrophage colony-stimulating factor (GM-CSF) and/or interleukin-3 (IL-3) on the globin program of adult human erythroid progenitors (BFUe) stimulated to terminal differentiation by erythropoietin under fetal bovine serum (FBS)-supplemented or FBS- deprived culture conditions. Fetal globin production by BFUe-derived erythroblasts was assessed at the protein and mRNA level and its cellular distribution was evaluated by immunofluorescence. Although hemoglobinization and maturation of BFUe-derived erythroblasts was by and large comparable in FBS-replete versus FBS-deprived cultures, the latter had significantly less (up to 20-fold) gamma-globin and gamma- globin mRNA levels. Reduced gamma-globin in serum-deprived cultures was also reflected by a smaller proportion of erythroblasts with detectable gamma-globin by immunofluorescence. Erythroid bursts induced by either GM-CSF or IL-3 produced similar levels of gamma-globin both in FBS- supplemented and in FBS-deprived cultures. These results, obtained even in cultures of highly enriched BFUe, suggest that GM-CSF and IL-3, although they significantly increase the number and size of erythroid bursts, do not by themselves exert a direct influence on the level of fetal globin synthesis. By contrast, factor(s) present in FBS appear to exert a dominant influence on fetal globin synthesis in vitro. Although FBS-deprived conditions appear to largely abrogate the in vitro activation of fetal hemoglobin (Hb F) in normal samples, they do support increased Hb F production in samples from patients with hereditary persistence of fetal hemoglobin or from cord blood.

scholarly journals Elucidation of the Interaction between Flavan-3-ols and Bovine Serum Albumin and Its Effect on Their In-Vitro Cytotoxicity

Molecules ◽  
2019 ◽  
Vol 24 (20) ◽  
pp. 3667
Author(s):  
Yasuyuki Fujii ◽  
Yoshitomo Suhara ◽  
Yusuke Sukikara ◽  
Tomohiro Teshima ◽  
Yoshihisa Hirota ◽  
...  
Keyword(s):  
Cell Culture ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
In Vitro Cytotoxicity ◽  
Binding Energies ◽  
Docking Simulations ◽  
Fetal Bovine

Flavan-3-ols (FLs), specifically catechin and its oligomer B-type procyanidins, are suggested to potently bind to bovine serum albumin (BSA). We examined the interaction between BSA and FLs by fluorescence quenching and found the following order of binding activities to BSA: cinnamtannin A2 (A2; tetramer) > procyanidin C1 (C1; trimer) ≈ procyanidin B2 (B2, dimer) > (−)epicatechin (EC, monomer). Docking simulations between BSA and each compound at the binding site showed that the calculated binding energies were consistent with the results of our experimental assay. FLs exerted cytotoxicity at 1000 μg/mL in F11 cell culture with fetal bovine serum containing BSA. In culture containing serum-free medium, FLs exhibited significant cell proliferation at 10−4 μg/mL and cytotoxicity was observed at concentrations greater than 10 μg/mL. Results of this study suggest that interactions between polyphenols and BSA should be taken into account when evaluating procyanidin in an in vitro cell culture system.

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