Reversible change in the fibroblast lysosomal enzyme dipeptidyl aminopeptidase-1 (cathepsin c) related to the commercial source of fetal bovine serum in the culture medium

In Vitro ◽  
1985 ◽  
Vol 21 (6) ◽  
pp. 340-346 ◽  
Author(s):  
Michael J. Doughty ◽  
Michael H. Davis ◽  
Eric Gruenstein
Keyword(s):  
Lysosomal Enzyme ◽  
Bovine Serum ◽  
Culture Medium ◽  
Fetal Bovine Serum ◽  
Reversible Change ◽  
Cathepsin C ◽  
Fetal Bovine ◽  
Commercial Source ◽  
Dipeptidyl Aminopeptidase

Fetal bovine serum induces changes in fatty acid composition ofTrypanosoma cruziphosphoinositides

1995 ◽  
Vol 41 (10) ◽  
pp. 951-954 ◽  
Author(s):  
G. Racagni ◽  
M. G. de Lema ◽  
G. Hernández ◽  
E. E. Machado-Domenech
Keyword(s):  
Fatty Acids ◽  
Fatty Acid Composition ◽  
Fatty Acid ◽  
Trypanosoma Cruzi ◽  
Acid Composition ◽  
Bovine Serum ◽  
Culture Medium ◽  
Culture Media ◽  
Fetal Bovine Serum ◽  
Fetal Bovine

Fetal bovine serum (FBS) is a necessary constituent of the culture media employed to foster the growth of Trypanosoma cruzi epimastigote forms. In different laboratories, the serum is used at final concentrations of 5 or 10%. We have normally supplemented the complex medium with 10% FBS. Under this condition we have described the fatty acid composition of the total lipids and of the phosphoinositide fractions. Additionally, we have reported the increase of polyphosphoinositides and phosphatidic acid after cholinergic stimulation. Since further attempts to reproduce these results with 5% FBS in the culture medium were not successful, the effect of the FBS concentration on the fatty acid composition of phospholipids from the T. cruzi epimastigote forms was thoroughly examined. This work showed that when the FBS concentration supplementing the culture medium was reduced from 10 to 5%, the fatty acid composition of the phosphoinositides was altered while the other major phospholipids were not significantly affected. The most relevant result was the decrease in the content of linoleic acid (18:2) and the increase of palmitoleic acid (16:1) in phosphatidylinositol 4,5-bisphosphate. Phosphatidylinositol (PI) and phosphatidylinositol phosphate also exhibited similar changes in the same fatty acids. The C2fatty acid composition of the phosphoinositides, under the same conditions, is also reported here for the first time.Key words: Trypanosoma cruzi, fatty acids, phosphoinositides, fetal bovine serum, phospholipids.

scholarly journals Platelet-Rich Fibrin Extract: A Promising Fetal Bovine Serum Alternative in Explant Cultures of Human Periosteal Sheets for Regenerative Therapy

2019 ◽  
Vol 20 (5) ◽  
pp. 1053 ◽  
Author(s):  
Tomoyuki Kawase ◽  
Masaki Nagata ◽  
Kazuhiro Okuda ◽  
Takashi Ushiki ◽  
Yoko Fujimoto ◽  
...  
Keyword(s):  
Bovine Serum ◽  
Culture Medium ◽  
Fetal Bovine Serum ◽  
Antigen Expression ◽  
Explant Culture ◽  
The Novel ◽  
Regenerative Therapy ◽  
Platelet Rich Fibrin ◽  
Alp Activity ◽  
Fetal Bovine

In 2004, we developed autologous periosteal sheets for the treatment of periodontal bone defects. This regenerative therapy has successfully regenerated periodontal bone and augmented alveolar ridge for implant placement. However, the necessity for 6-week culture is a limitation. Here, we examined the applicability of a human platelet-rich fibrin extract (PRFext) as an alternative to fetal bovine serum (FBS) for the explant culture of periosteal sheets in a novel culture medium (MSC-PCM) originally developed for maintaining mesenchymal stem cells. Small periosteum tissue segments were expanded in MSC-PCM + 2% PRFext for 4 weeks, and the resulting periosteal sheets were compared with those prepared by the conventional method using Medium199 + 10% FBS for their growth rate, cell multilayer formation, alkaline phosphatase (ALP) activity, and surface antigen expression (CD73, CD90, and CD105). Periosteal sheets grew faster in the novel culture medium than in the conventional medium. However, assessment of cell shape and ALP activity revealed that the periosteal cells growing in the novel medium were relatively immature. These findings suggest that the novel culture medium featuring PRFext offers advantages by shortening the culture period and excluding possible risks associated with xeno-factors without negatively altering the activity of periosteal sheets.

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.

A combination of bovine serum albumin with insulin–transferrin–sodium selenite and/or epidermal growth factor as alternatives to fetal bovine serum in culture medium improves bovine embryo quality and trophoblast invasion by induction of matrix metalloproteinases

2019 ◽  
Vol 31 (2) ◽  
pp. 333 ◽  
Author(s):  
Ayman Mesalam ◽  
Kyeong-Lim Lee ◽  
Imran Khan ◽  
M. M. R. Chowdhury ◽  
Shimin Zhang ◽  
...  
Keyword(s):  
Bovine Serum ◽  
Sodium Selenite ◽  
Culture Medium ◽  
Fetal Bovine Serum ◽  
Cell Number ◽  
Matrix Metalloproteinase 2 ◽  
Bovine Embryo ◽  
Epidermal Growth ◽  
Fetal Bovine ◽  
Long Chain

This study investigated the use of bovine serum albumin (BSA) plus insulin–transferrin–sodium selenite (ITS) and/or epidermal growth factor (EGF) as alternatives to fetal bovine serum (FBS) in embryo culture medium. The developmental ability and quality of bovine embryos were determined by assessing their cell number, lipid content, gene expression and cryotolerance, as well as the invasion ability of trophoblasts. The percentage of embryos that underwent cleavage and formed a blastocyst was higher (P<0.01) in medium containing ITS plus EGF and BSA than in medium containing FBS. Culture with ITS plus EGF and BSA also increased the hatching ability of blastocysts and the total cell number per blastocyst. Furthermore, the beneficial effects of BAS plus ITS and EGF on embryos were associated with a significantly reduced intracellular lipid content, which increased their cryotolerance. An invasion assay confirmed that culture with ITS plus EGF and BSA significantly improved the invasion ability of trophoblasts. Real-time quantitative polymerase chain reaction analysis showed that the mRNA levels of matrix metalloproteinase-2 (MMP2) and MMP9, acyl-CoA synthetase long-chain family member 3, acyl-coenzyme A dehydrogenase long-chain and hydroxymethylglutaryl-CoA reductase significantly increased upon culture with ITS plus EGF and BSA. Moreover, protein expression levels of matrix metalloproteinase-2 and -9 increased (P<0.01) in medium supplemented with ITS plus EGF and BSA compared with medium supplemented with FBS. Taken together, these data suggest that supplementation of medium with ITS plus EGF and BSA improves invitro bovine embryo production, cryotolerance and invasion ability of trophoblasts.

Adaptation of an Aedes aegypti cell line to hemolymph-free culture medium

1969 ◽  
Vol 15 (10) ◽  
pp. 1197-1200 ◽  
Author(s):  
S. S. Sohi
Keyword(s):  
Aedes Aegypti ◽  
Bombyx Mori ◽  
Cell Line ◽  
Bovine Serum ◽  
Culture Medium ◽  
Fetal Bovine Serum ◽  
Free Medium ◽  
Antheraea Pernyi ◽  
Fetal Bovine ◽  
In The Beginning

Aedes aegypti cells (Grace's line) growing in a medium containing 5% Antheraea pernyi hemolymph (APH) and 5% fetal bovine serum (FBS) were adapted to a hemolymph-free medium. The quantity of APH was gradually substituted with an equal quantity of FBS over a 2-month period. The cells grew very slowly in the hemolymph-free medium in the beginning. However, they became adapted to it in another 8 months; after that they grew as well as the cells in medium supplemented with hemolymph.Also, these cells grew as well in medium containing Bombyx mori hemolymph as in medium enriched with APH.

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.

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