In vitro cultivation of hemocytes of Malacosoma disstria Hübner (Lepidoptera: Lasiocampidae)

1971 ◽  
Vol 49 (10) ◽  
pp. 1355-1358 ◽  
Author(s):  
S. S. Sohi
Keyword(s):  
Tissue Culture ◽  
Culture Medium ◽  
Fetal Bovine Serum ◽  
Primary Cultures ◽  
Malacosoma Disstria ◽  
In Vitro Cultivation ◽  
Original Source ◽  
Insect Tissue Culture ◽  
Fetal Bovine

Prolonged culturing of the hemocytes of Malacosoma disstria has been accomplished using Grace's insect tissue culture medium supplemented with fetal bovine serum (5%) and Bombyx mori hemolymph (3%). The cultures started to grow after 3–6 months. These cells have now been in vitro for over 16 months, and have been subcultured 35 times. Three types of cells were present in primary cultures, but only one type, prohemocytes, persisted and grew after subculturing. The M. disstria larvae that were used as the original source of hemocytes were naturally infected with the microsporidian Glugea disstriae. The microsporidian also grew in [he cell cultures, and the cells are still infected.

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.

Soy protein isolate: A substitute of fetal bovine serum for the in vitro cultivation of Leishmania donovani

2017 ◽  
Author(s):  
Arushdeep Sidana ◽  
Afroz Alam ◽  
Umar Farooq
Keyword(s):  
Soy Protein ◽  
Bovine Serum ◽  
Culture Medium ◽  
Leishmania Donovani ◽  
Fetal Bovine Serum ◽  
Soy Protein Isolate ◽  
Protein Isolate ◽  
In Vitro Cultivation ◽  
Fetal Bovine

Fetal Bovine Serum (FBS) is an expensive source of macronutrients which are required for proper nutrition of Leishmania parasite in the culture medium. An alternative, cost effective source of macronutrients which can replace the use of FBS in tissue culture medium is required. The potential of Soy Protein Isolate (SPI) to replace FBS in RPMI-1640 medium for the in vitro cultivation of Leishmania donovani was evaluated. Commercially available SPI powder was used in RPMI-1640 medium as a substitute of FBS to cultivate L. donovani promastigotes. The growth, multiplication and morphology of cultivated parasites was observed in conventional RPMI-1640 with 10% FBS (v/v) and RPMI-1640 containing 10% SPI (v/v) by using light microscopy, measurement of absorbance and cell counting. The growth of Leishmania promastigotes in the medium containing 10% SPI was slower in initial phase; however, the parasites were morphologically larger as compared to those in RPMI-1640 medium containing 10% FBS. Cell count in the SPI-containing RPMI-1640 medium was 2.3 × 108 cells/ml whereas it was 1.9 × 107 cells/ml in RPMI-1640 with 10% FBS. This study concludes that RPMI-1640 may be supplemented with SPI instead of FBS for the in vitro cultivation of Leishmania donovani promastigotes to decrease the culture maintenance cost in developing countries.

134 EFFECT OF REPLACING FETAL BOVINE SERUM BY DIFFERENT GROWTH FACTORS IN THE PRE-IMPLANTATION DEVELOPMENT OF BOVINE EMBRYOS GENERATED BY IN VITRO FERTILIZATION

2014 ◽  
Vol 26 (1) ◽  
pp. 180
Author(s):  
R. Felmer ◽  
T. Vargas ◽  
R. Sanchez ◽  
M. E. Arias
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Culture Medium ◽  
Fetal Bovine Serum ◽  
Fibroblast Growth Factor 2 ◽  
Bovine Embryos ◽  
Epidermal Growth ◽  
Fetal Bovine ◽  
Defined Culture

Different culture systems have been studied that support pre-implantation development of bovine embryos up to the blastocyst stage. However, the use of chemically defined culture systems has been less studied. The objective of the present study was to evaluate the effect, in the developmental potential of in vitro-produced bovine embryos, of replacing fetal bovine serum (FBS) by different growth factors in the maturation and embryo culture media. In experiment 1, oocytes collected by aspiration of ovaries from a local slaughterhouse were matured in standard TCM-199 culture medium at 38.5°C, 5% CO2, and saturation humidity. The effect of insulin-like growth factor 1 (100 ng mL–1), epidermal growth factor (10 ng mL–1), and fibroblast growth factor 2 (500 ng mL–1) was evaluated at 24 h by the presence of a polar body after removal of cumulus-oocyte complexes. In experiment 2, oocytes matured in vitro in the presence of FBS were fertilized by co-incubation with commercial sperm (mL) for 18 h in standard fertilization medium (Fert-TALP). The presumptive zygotes were denuded and randomly allocated in a chemically defined culture medium based on KSOM supplemented with polyvinyl alcohol (PVA), fructose, and each of the growth factors listed previously. Undefined cultured medium was based on KSOM supplemented with 5% FBS. Embryos were cultured at 38.5°C in a mixture of gases and saturation humidity. Cleavage and blastocyst rates were recorded on Days 3 and 7, respectively. Analysis of variance was used to test for statistically significant differences between groups (P < 0.05) using Stat Graphics Plus 2 Software. In cases where statistically significant differences were observed, a multiple comparison test was run using Tukey's test. In experiment 1, a similar maturation rate was observed in all treatments relative to the undefined maturation medium (range = 88–91%). In experiment 2, no differences were observed in the cleavage (79, 87, 85, and 85%) and the blastocyst rates (24, 25, 26, and 30%) for the epidermal growth factor, insulin-like growth factor 1, fibroblast growth factor 2, and FBS treatments, respectively. In conclusion, we demonstrated that maturation of bovine oocytes can be achieved in chemically defined conditions by replacing FBS by each of the growth factors evaluated herein. Furthermore, chemically defined KSOM medium supplemented by any of these growth factors can generate a similar rate of blastocyst than the undefined medium containing FBS. Analyses are under way to evaluate the effect of completely defined culture conditions (maturation and embryo culture) on the pre-implantation development of embryos produced in the presence of these growth factors.

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&amp;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.

Silicate and Calcium Ions Releasing Biomaterials for Bone Reconstruction

2011 ◽  
Vol 493-494 ◽  
pp. 561-565
Author(s):  
Jin Nakamura ◽  
Akiko Obata ◽  
Julian R. Jones ◽  
Toshihiro Kasuga
Keyword(s):  
Calcium Ions ◽  
Culture Medium ◽  
Fetal Bovine Serum ◽  
Poly Lactic Acid ◽  
Cell Culture Medium ◽  
A Cell ◽  
Fetal Bovine ◽  
Mouse Osteoblast ◽  
Phosphate Deposits

Siloxane-containing vaterite (SiV) / poly (lactic acid) hybrid (SiPVH) beads with the releasability of silicate and calcium ions were prepared with an electrospraying method. According to the increase in the silicon content of the SiV, the amount of silicate ion released from the resulting beads also increased. When the beads were soaked in a cell culture medium, proteins derived from fetal bovine serum were adsorbed on their surfaces. Cell adhesion tests were also performed on the beads with using mouse osteoblast-like cell line (MC3T3-E1) in vitro. After 5 days of culturing, the cells adhered and spread well to cover the surface of the beads. In the localized area, agglomerated cells were observed to combine with cauliflower-shaped calcium phosphate deposits.

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: &gt;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: &lt;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&lt;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&lt;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&lt;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&lt;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.

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