Growth and development of rabbit oocytes in vitro: Effect of fetal bovine serum concentration on culture medium

2012 ◽  
Vol 78 (5) ◽  
pp. 1040-1047 ◽  
Author(s):  
H. Sugimoto ◽  
Y. Kida ◽  
Y. Miyamoto ◽  
K. Kitada ◽  
K. Matsumoto ◽  
...  
Keyword(s):  
Serum Concentration ◽  
Bovine Serum ◽  
Growth And Development ◽  
Culture Medium ◽  
Fetal Bovine Serum ◽  
Fetal Bovine Serum Concentration ◽  
Fetal Bovine ◽  
Vitro Effect

scholarly journals INVESTIGATING COOLING RATE AND FETAL BOVINE SERUM CONCENTRATION ON SURVIVAL AND STEMNESS OF MESENCHYMAL STEM CELLS AFTER CRYOPRESERVATION

2009 ◽  
Vol 12 (9) ◽  
pp. 12-22
Author(s):  
Phuc Van Pham ◽  
Tam Thanh Nguyen ◽  
Nhung Thi Hong Vuong ◽  
Tuyet Thi Bach Duong ◽  
Ngoc Kim Phan
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Serum Concentration ◽  
Cord Blood ◽  
Umbilical Cord Blood ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Fetal Bovine Serum Concentration ◽  
Fetal Bovine ◽  
Different Sources

Mesenchymal stem cells (MSCs) can be derived from many different sources. Umbilical cord blood is a rich source of MSCs. The cryopreservation of MSCs that MSCs are still alive and differentiate into many different kinds of functional cells is very important. The aims of this research are to identify ratio of alive and dead cells as well as stemness of them after thaw. The results showed that the stemness was not affected by cryopreservative protocols or media. All cells being alive after thaw could form colonies and differentiate into adipocytes and osteoblasts. Ratio of alive and dead cells was affected very much by cryopreservative protocols and media.

117 Kit ligand enhances growth and nuclear maturation of buffalo oocytes in vitro

2019 ◽  
Vol 31 (1) ◽  
pp. 184
Author(s):  
M. N. Islam ◽  
M. H. Alam ◽  
A. Khatun ◽  
M. A. Hashem ◽  
M. Moniruzzaman
Keyword(s):  
Serum Sodium ◽  
Bovine Serum ◽  
Culture Medium ◽  
Fetal Bovine Serum ◽  
Survival Rates ◽  
Sodium Pyruvate ◽  
Antral Follicles ◽  
Kit Ligand ◽  
Fetal Bovine

This study aimed to investigate the effect of Kit ligand (KL), a growth factor that regulates folliculogenesis in mammalian ovaries, on growth of buffalo oocytes in early antral follicles in vitro. Cumulus-oocyte complexes were dissected from early antral follicles (1mm) of slaughtered buffaloes and cultured in Dulbecco’s minimum essential medium supplemented with fetal bovine serum, sodium pyruvate, gentamycin, hypoxanthine, dexamethasone, cysteine, polyvinylpyrolidione, l-ascorbic acid, oestradiol-17β, and androstenedione in a 96-well culture plate at 38.5°C under an atmosphere of 5% CO2 in air for 6 days. The culture medium was supplemented with 0, 50, and 100 ng/mL KL (recombinant human SCF, Cat. No. H8416, R&D Systems, Minneapolis, MN, USA). Sixty oocytes were cultured in each group with 6 replications. In vitro-grown oocytes were cultured for maturation in tissue culture medium-199 supplemented with 5% fetal bovine serum, sodium pyruvate, gentamycin, and 100 ng/mL FSH at 38.5°C for 24h under an atmosphere of 5% CO2 in air. The oocytes were then stained with aceto-orcein and examined under a differential interference contrast microscope. Data were analysed using SAS/STAT version 9.1.3 for Windows (SAS Institute Inc., Cary, NC, USA) by one-way ANOVA and means compared with Tukey’s HSD test. The mean diameter of oocytes measured at the time of seeding on the culture substrate was 100.6±0.4μm (n=180). After 6 days of culture, the diameters of oocytes increased to 110.8±0.5, 114.0±0.5, and 115.0±0.6µm in 0, 50, and 100 ng/mL KL-treated groups, respectively. The survival rates were 60.0±6, 81.2±1.2, and 92.0±4.9% in 0, 50, and 100 ng/mL KL-supplemented oocytes at Day 6. Moreover, KL pretreatment enhanced maturation of buffalo oocytes dose dependently. A small proportion of oocytes (8.4%) treated with 50 ng/mL KL reached the MII stage. This number increased to 25% when oocytes were treated with 100 ng/mL KL. These results show that KL enhances growth, viability, and meiotic progression of buffalo oocytes in vitro.

scholarly journals 2POSTHATCHING DEVELOPMENT SYSTEM: A NOVEL IN VITRO CULTURE OF BOVINE EMBRYOS

2004 ◽  
Vol 16 (2) ◽  
pp. 123 ◽  
Author(s):  
D.O. Brandão ◽  
G. Vajta ◽  
P. Maddox-Hyttel ◽  
D. Stringfellow ◽  
P. Lövendahl ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Bovine Serum ◽  
Culture System ◽  
Culture Medium ◽  
Inner Cell Mass ◽  
Fetal Bovine Serum ◽  
Cell Mass ◽  
Embryo Morphology ◽  
Fetal Bovine

Although high blastocyst rates can be achieved in somatic cell nuclear transfer, abortions and developmental abnormalities still hamper advancement. Reliable and practical methods to evaluate early embryonic development and differentiation are required to understand and overcome the problem. Our aim was to establish an in vitro culture system for monitoring posthatching development (PHD). Slaughterhouse-derived bovine oocytes were matured in vitro, fertilized (Day 0) and cultured (Holm et al., 1999, Theriogenology, 52, 683–700). On Day 8, degenerated embryos were removed from each well and 400L of modified culture medium (SOFaaci plus 0.5% glucose and 10% fetal bovine serum) were added. At Day 11, hatched blastocysts were selected by scoring them as Quality 1 (Q1: >1.0mm, clear trophoblast, compact inner cell mass), Quality 2 (Q2: 0.5mm, dark spots in the trophoblast, less compact inner cell mass), or Quality 3 (Q3: <0.5mm, many dark spots in the trophoblast, spread inner cell mass). The resulting 304 blastocysts in 12 replicates were then loaded into 15mm×1.2 gel tunnels of 2.4% agarose in PBS, supplemented with either 5% (Agar5) or 10% (Agar10) fetal bovine serum, covered with the modified culture medium, and then incubated at 38.5°C in 5% CO2, 5% O2, 90% N2. Embryo morphology and length were evaluated using a stereomicroscope on Days 12, 13, 14 and 15. On Day 14, 75 embryos were removed, biopsed (1mm) for sex determination of each embryo, and processed for light and transmission electron microscopy. Qualitative and quantitative data were analyzed by χ2 test and GLM procedure of SAS, respectively, with P level of 0.05. A total of 170 embryos (56% of total) initiated elongation. This percentage was higher (LSmeansSD, n=12; P<0.05) in Agar10 v. Agar5 in both Q1 (889 v. 637), Q2 (667 v. 485) and Q3 embryos (529 v. 278). Mean embryo length (mm; LSmeansSEM) on Day 13 was higher (P<0.05) in Q1 (2.10.2, n=49) and Q2 (1.71.4, n=98) than Q3 (1.20.3, n=23). On Day 14, Q1 embryos (3.50.2) were longer (P<0.01) than Q2 and Q3 embryos (2.70.1 and 2.00.3). On Day 15, Q1, Q2 and Q3 embryos (4.40.5, n=24, 4.00.3, n=45 and 2.90.6, n=14, respectively) had similar length, probably influenced by the low number of Q3 embryos. The percentage of males was higher (P<0.001) in Q1 (95%; n=40), but similar in Q2 (39%; n=26) and Q3 (71%; n=7). Light microscopy confirmed hypoblast and epiblast formation. Ultrastructural analysis revealed that the latter had penetrated the trophoblast (Rauber’s layer), forming an embryonic disc including many degenerative cells. In conclusion, this culture system represents the first model for rapid growth, elongation, and initial differentiation of bovine posthatching embryos.

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.

Fetal Bovine Serum Concentration Affects Δ9 Desaturase Activity of Trypanosoma cruzi

Lipids ◽  
2010 ◽  
Vol 45 (3) ◽  
pp. 275-283 ◽  
Author(s):  
Ana L. Villasuso ◽  
Patricio Romero ◽  
Mariela Woelke ◽  
Patricia Moyano ◽  
Estela Machado ◽  
...  
Keyword(s):  
Serum Concentration ◽  
Trypanosoma Cruzi ◽  
Bovine Serum ◽  
Desaturase Activity ◽  
Fetal Bovine Serum ◽  
Fetal Bovine Serum Concentration ◽  
Δ9 Desaturase ◽  
Fetal Bovine

IN VITRO RAT ISLET CULTURE WITH SILK PROTEIN SERICIN, A NEW CULTURE MEDIUM INSTEAD OF FETAL BOVINE SERUM.

2004 ◽  
Vol 78 ◽  
pp. 540-541 ◽  
Author(s):  
M Morikawa ◽  
T Kimura ◽  
M Murakami ◽  
K Katayama ◽  
S Terada ◽  
...  
Keyword(s):  
Bovine Serum ◽  
Culture Medium ◽  
Fetal Bovine Serum ◽  
Silk Protein ◽  
Islet Culture ◽  
Fetal Bovine

scholarly journals TINGKAT PERKEMBANGAN AWAL EMBRIO SAPI IN VITRO MENGGUNAKAN MEDIA TUNGGAL BERBAHAN DASAR TISSUE CULTURE MEDIUM (TCM) 199

2013 ◽  
Vol 7 (2) ◽  
Author(s):  
Mohamad Agus Setiadi ◽  
Ni Wayan Kurniani Karja
Keyword(s):  
Tissue Culture ◽  
Bovine Serum Albumin ◽  
Chorionic Gonadotropin ◽  
Bovine Serum ◽  
Culture Medium ◽  
Fetal Bovine Serum ◽  
Tissue Culture Medium ◽  
Serum Gonadotropin ◽  
Fetal Bovine

Penelitian ini bertujuan mengetahui kemampuan perkembangan awal embrio sapi in vitro menggunakan media tunggal untuk maturasi, fertilisasi, dan kultur berbahan dasar tissue culture medium (TCM) 199. Oosit sapi dikumpulkan dari rumah potong hewan dengan teknik aspirasi dan diklasifikasikan berdasarkan kekompakan sel kumulus dan sitoplasma yang homogen. Oosit dimaturasi pada medium TCM 199 yang disuplementasi dengan 10 IU/ml pregnant mare’s serum gonadotropin (PMSG), 10 IU/ml human chorionic gonadotropin (hCG), dan 10% fetal bovine serum (FBS), dilakukan selama 24 jam pada inkubator 5% CO2, 39 C. Fertilisasi dilakukan pada dua media yang berbeda yaitu media rutin fertilisasi dan media berbahan dasar TCM 199 dengan suplemen bovine serum albumin (BSA) dan heparin. Setelah fertilisasi, kumulus sel dihilangkan (denudasi), kemudian dikultur pada media TCM 199 yang disuplementasi dengan asam amino esensial dan non-esensial serta 10% FBS selama 3 hari. Hasil penelitian menunjukkan tingkat maturasi oosit pada sistem yang digunakan mampu mendukung 81,5% oosit mencapai tahap metafase II (M-II). Tingkat pembelahan embrio lebih tinggi pada media rutin dibandingkan dengan media TCM 199 yakni masing-masing 44,4 dan 23,2%. Jumlah embrio tahap 4-8 sel pada kedua perlakuan tidak berbeda nyata. Dapat disimpulkan media tunggal berbasis TCM dapat digunakan untuk produksi embrio in vitro.

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