Influence of calcium on the differentiation and growth of aggregated organs in Sphaerostilbe repens

1978 ◽  
Vol 24 (9) ◽  
pp. 1039-1047 ◽  
Author(s):  
B. Botton
Keyword(s):  
Culture Medium ◽  
Defined Medium ◽  
Vegetative Mycelium ◽  
Sphaerostilbe Repens

In the ascomycete Sphaerostilbe repens the vegetative mycelium aggregated into structures composed of aerial coremia and rhizomorphs immersed in the culture medium. The morphology and growth of the cultures were examined on a defined medium with and without calcium supplementation.Initiation and growth of aggregated organs occurred only if calcium was present continuously in the culture medium. On calcium-deficient media the fungus produced only vegetative mycelium which retained its ability to aggregate in the presence of the cation. Small concentrations of calcium (less than 1 ppm) stimulated aggregation of the mycelium, but higher concentrations (about 200 ppm) were necessary to obtain optimum development of the aggregated organs. Strontium partially replaced calcium, but lithium did not, as a stimulator of aggregation.

scholarly journals Transcriptional Profiling of Porcine Blastocysts Produced In Vitro in a Chemically Defined Culture Medium

Animals ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1414
Author(s):  
Josep M. Cambra ◽  
Emilio A. Martinez ◽  
Heriberto Rodriguez-Martinez ◽  
Maria A. Gil ◽  
Cristina Cuello
Keyword(s):  
Culture Medium ◽  
Embryo Quality ◽  
Transcriptional Profiling ◽  
Defined Medium ◽  
Platelet Factor ◽  
Defined Media ◽  
Defined Culture ◽  
The Impact

The development of chemically defined media is a growing trend in in vitro embryo production (IVP). Recently, traditional undefined culture medium with bovine serum albumin (BSA) has been successfully replaced by a chemically defined medium using substances with embryotrophic properties such as platelet factor 4 (PF4). Although the use of this medium sustains IVP, the impact of defined media on the embryonic transcriptome has not been fully elucidated. This study analyzed the transcriptome of porcine IVP blastocysts, cultured in defined (PF4 group) and undefined media (BSA group) by microarrays. In vivo-derived blastocysts (IVV group) were used as a standard of maximum embryo quality. The results showed no differentially expressed genes (DEG) between the PF4 and BSA groups. However, a total of 2780 and 2577 DEGs were detected when comparing the PF4 or the BSA group with the IVV group, respectively. Most of these genes were common in both in vitro groups (2132) and present in some enriched pathways, such as cell cycle, lysosome and/or metabolic pathways. These results show that IVP conditions strongly affect embryo transcriptome and that the defined culture medium with PF4 is a guaranteed replacement for traditional culture with BSA.

Feeding of Boophilus microplus larvae on a partially defined medium through thin slices of cattle skin

Parasitology ◽  
1975 ◽  
Vol 70 (2) ◽  
pp. 243-254 ◽  
Author(s):  
D. H. Kemp ◽  
D. Koudstaal ◽  
J. A. Roberts ◽  
J. D. Kerr
Keyword(s):  
Bovine Serum ◽  
Freeze Drying ◽  
Culture Medium ◽  
Culture Method ◽  
Defined Medium ◽  
Boophilus Microplus ◽  
Cattle Tick ◽  
Haemaphysalis Longicornis ◽  
Freeze Dried ◽  
Thin Slices

Larvae of the cattle tick Boophilus microplus will attach to thin (0·3–0·5 mm) slices of cattle skin and engorge on a partially denned medium at 35 °C. Forty-seven to 83% of the larvae had engorged by 8 days, and 51–71% of these moulted to nymphs. Tissue culture medium alone allowed little growth unless supplemented with dialysed, freeze dried bovine serum (7%, w/v). This medium could be further defined by substituting purified bovine serum albumin (Cohn fraction V) for the dialysed bovine serum. In one experiment, nymphs of Haemaphysalis longicornis engorged and later moulted to adults. Skin slices were used fresh or after freeze-drying and storing at − 25 °C. The possible uses of the culture method are discussed.

scholarly journals Schistosomula of Schistosoma mansoni clear concanavalin A from their surface by sloughing.

1982 ◽  
Vol 94 (2) ◽  
pp. 355-362 ◽  
Author(s):  
J C Samuelson ◽  
J P Caulfield ◽  
J R David
Keyword(s):  
Schistosoma Mansoni ◽  
Concanavalin A ◽  
Binding Sites ◽  
Culture Medium ◽  
Culture Media ◽  
Lectin Binding ◽  
Defined Medium ◽  
Con A ◽  
Sds Page ◽  
Lectin Binding Sites

The lectin concanavalin A (Con A) was used as a model probe to study the behavior of molecules bound to the surface of recently transformed schistosomula of Schistosoma mansoni. Con A binding was saturable (150-180 pg/organism) and specifically competed by alpha-methyl mannoside. Both FITC-Con A and 125-I-Con A were lost from the surface of schistosomula with a halftime of 8-10 h in culture in defined medium. A comparable decrease in the binding of Con A to schistosomula cultured and then labeled with the lectin indicated that the labeling procedure itself was not inducing the observed change. Internalization of Con A was not seen by either fluorescence microscopy or electron microscope radioautography. In addition, 70-80% of the radioactivity lost from the parasite was recoverable by TCA precipitation from the culture medium as intact Con A (27,000 mol wt on SDS PAGE). Thus, the mechanism of clearance of bound Con A from the surface of cultured schistosomula is apparently by sloughing of Con A molecules intact into the culture media and not by endocytosis and degradation. Con A binding sites, visualized with hemocyanin by scanning electron microscopy, appeared homogeneously distributed over the surface of schistosomula when organisms were labeled at 4 degree C or after fixation with glutaraldehyde. However, Con A and hemocyanin formed aggregates on the surface of schistosomula when labeling was performed at 37 degrees C, which suggests that lectin binding sites have lateral mobility within the plane of the membrane. These aggregates are likely independent of metabolism by the parasite because aggregation also occurs on the surface of organisms killed with azide.

scholarly journals The glycosulphatase of Trichoderma viride

1968 ◽  
Vol 108 (3) ◽  
pp. 393-399 ◽  
Author(s):  
A G Lloyd ◽  
P J Large ◽  
M Davies ◽  
A H Olavesen ◽  
K S Dodgson
Keyword(s):  
Culture Medium ◽  
Enzyme System ◽  
Trichoderma Viride ◽  
Defined Medium ◽  
Potassium Sulphate ◽  
Experimental Conditions ◽  
Lag Period

The growth of the mould Trichoderma viride on a defined medium containing either potassium d-glucose 6-O-sulphate or potassium d-galactose 6-O-sulphate as sole sources of both carbon and sulphur is marked by the production of an enzyme system capable of liberating inorganic SO42− ions from either of the sulphate esters. The enzyme is not produced when the organism is grown with glucose (or galactose) and potassium sulphate or with glucose and methionine as sole sources of carbon and sulphur. Experimental conditions are described whereby inorganic SO42− ions liberated from potassium glucose 6-O-sulphate by the growing mould appear in the culture medium after a constant lag period of 21–24hr. The enzyme has been shown to be a simple glycosulphatase that is active towards the 6-O-sulphate esters of d-glucose and d-galactose but not towards potassium glucose 3-O-sulphate. The properties of the crude glycosulphatase show the enzyme to be appreciably different from analogous molluscan enzymes that can degrade monosaccharide sulphate esters.

scholarly journals In vitro production of cattle blastocysts in chemically defined medium with or without insulin supplementation

1996 ◽  
Vol 5 (5) ◽  
pp. 509-514 ◽  
Author(s):  
Kristiina Bredbacka ◽  
Peter Bredbacka
Keyword(s):  
Bovine Serum Albumin ◽  
Culture Medium ◽  
Defined Medium ◽  
Blastocyst Stage ◽  
Chemically Defined Medium ◽  
In Vitro Production ◽  
Cell Numbers ◽  
Vitro Production ◽  
Bovine Oocytes

In this study we evaluated the use of a chemically defined medium in the production of blastocysts from bovine oocytes fertilized in vitro. As culture medium we used CRI-PVP, a modification of CRlaa medium with bovine serum albumin replaced by polyvinylpyrrolidone. After 168 h of culture (192 h after insemination) 8.7%, 10.5 and 12.8% of the cleaved embryos developed to the blastocyst stage in the presence of 0, 2 or 200 nM insulin, respectively. The supplementation of 200 nM insulin tended to increase cell numbers in morulae and blastocysts (P=0.10). It is concluded that CRI-PVP can be used as a chemically defined medium in the production of blastocysts from bovine 1-cell embryos. However, further modifications are needed, and the insulin concentrations used may be below the optimum for blastocyst production.

203 PI3-K PATHWAY IS ONE OF THE KEY REGULATORS OF STEROIDOGENESIS OF CUMULUS CELLS OF BOVINE CUMULUS–OOCYTE COMPLEXES MATURATED IN DEFINED MEDIUM IN THE PRESENCE OF FSH

2015 ◽  
Vol 27 (1) ◽  
pp. 192
Author(s):  
D. Kaiser de Souza ◽  
L. P. Salles ◽  
R. Camargo ◽  
B. Dolabela de Lima ◽  
F. A. G. Torres ◽  
...  
Keyword(s):  
Gene Expression ◽  
Culture Medium ◽  
Statistical Significance ◽  
Cumulus Cells ◽  
Defined Medium ◽  
Steroid Production ◽  
Maturation Medium ◽  
Bonferroni Test ◽  
Group Student

The aim of the present study was to access the function of PI3-K pathway tested by the use of its inhibitor, LY294002, in maturation medium of bovine cumulus-oocyte complexes (COC) in steroid concentration in the medium, key enzymes of steroidogenic pathway and gonadotrophin receptors of cumulus cells. This study was performed in defined medium without serum and albumin (MIV B) in absence or presence of 10 ng mL–1 of FSH. Androstenedione 10–7 M was used as a precursor of steroidogenesis. Bovine COC (n = 35–40/well) collected from ovaries obtained at abattoirs of Brasilia (DF, Brazil) were cultivated in 400 μL of medium MIV B; MIV B + 100 μmol mL–1 of LY294002; MIV B added to 10 ng mL–1 of FSH; or MIV B added to 10 ng mL–1 of FSH + 100 μmol mL–1 of LY294002 for 22 to 24 h. After culture, COC were mechanically denuded and cumulus cells from 20 COC were isolated by centrifugation to determine the gene expression of LHR, FSHR, CYP11A1, CYP19A1, and HSD17B1 by PCR real time. Cumulus cells of immature COC were also collected and analysed as the calibrator group. Student–Newman–Keuls was performed as statistical test. The culture medium was collected after culture to determine progesterone and 17β-oestradiol concentration by quimioluminescence method and to calculate E2/P4 ratio. Two-way ANOVA, followed by Bonferroni test, and t-test were performed to determine the statistical significance. The MIVB enhanced LHR, FSHR, CYP11A1, CYP19A1, and HSD17B1 and LY294002 inhibited the expression of all genes (P < 0.05). MIVB + FSH decreased the expression of all genes (P < 0.05) except CYP11A1. LY294002 in the presence of 10 ng mL–1 of FSH did not affect the gene expression of FSHR, CYP11A1, and HSD17B1; however, it increased expression of LHR and CYP19A1. Oestradiol and progesterone production was increased by supplementation of FSH in MIVB (P < 0.05), but the E2/P4 ratio did not differ between treatments. LY294002 decreased oestradiol and E2/P4 ratio in absence or presence of FSH (P < 0.05), but did not alter progesterone concentration in MIVB+FSH. The inhibitor of PI3-K decreased the expression of steroidogenic proteins and steroid production in absence of FSH. The supplementation of FSH increased steroid production and decreased gene expression of steroidogenic enzymes, except CYP11A1. The inhibition of PI3-K in presence of FSH increases LHR and CYP19A1 expression. This fact suggests a strong role of the PI3-K pathway in the regulation of LHR and CYP19A1 expression.The authors thank FAP-DF (193.000.577/2009), CNPq, CAPES, Sabin, and Ponte Alta abattoir, Brasilia.

Preliminary Experiments with a Defined Culture Medium for Larval Fasciola hepatica

1973 ◽  
Vol 47 (2) ◽  
pp. 181-189 ◽  
Author(s):  
R. S. V. Pullin
Keyword(s):  
Amino Acids ◽  
Culture Medium ◽  
Fasciola Hepatica ◽  
Dry Weight ◽  
Defined Medium ◽  
Inorganic Salts ◽  
Final Maturation ◽  
Defined Culture ◽  
Stock Solutions

A defined medium is described as a basis for in vitro culture work with larval Fasciola hepatica. This medium, termed BCM, can be quickly made up by using a system of stock solutions. BCM contains inorganic salts, glucose, amino acids, vitamins and antibiotics, but no lipid or proteins. Rediae can be dissected from infected snails for culture, but many appear to be contaminated with bacteria. Large rediae cannot survive in BCM but free immature cercariae can complete their final maturation in vitro. This final maturation, from the 30th to the 35th day after miracidial penetration of donor snails, includes tail growth and appearance of body pigmentation. Cercariae matured in vitro encyst successfully when transferred from BCM to water. Small rediae survive in BCM for 5 days, but show no growth or development measured as dry weight and total nitrogen.

A Chemically Defined Medium for Disinfectant Testing

1964 ◽  
Vol 47 (3) ◽  
pp. 539-539
Author(s):  
Michael J Pelczar
Keyword(s):  
Culture Medium ◽  
Defined Medium ◽  
Chemically Defined Medium ◽  
Testing Procedures ◽  
Collaborative Testing ◽  
Satisfactory Performance

Abstract A synthetic broth has been developed as a culture medium for use in disinfectant testing procedures, 5.001— 5.005. Collaborative testing of the Wright- Mundy medium revealed satisfactory performance, and the medium is recommended for adoption as official, first action, as an alternative medium for disinfectant testing.

scholarly journals Factors influencing the release of proteins by cultured schwann cells

1981 ◽  
Vol 91 (3) ◽  
pp. 666-672 ◽  
Author(s):  
DJ Carey ◽  
RP Bunge
Keyword(s):  
Schwann Cells ◽  
Culture Medium ◽  
Polyacrylamide Gel Electrophoresis ◽  
Qualitative Difference ◽  
Defined Medium ◽  
Normal Pattern ◽  
Fourfold Increase ◽  
Molecular Weights ◽  
Sds Page ◽  
Cultured Schwann Cells

Cultured rat schwann cells grown in association with sensory neurons when labeled with [(3)H]leucinem, [(3)H]glucosamine, or [(35)S]methionine release labeled polypeptides into the culture medium. Analysis by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) of the culture medium reveals a reproducible pattern of more than 20 polypeptides with molecular weights ranging from 15,000 to more than 250,000. Five major polypeptides (apparent molecular weights 225,000, 210,000, 90,000, 66,000, 50,000, and 40,000) account for approximately 40 percent of the leucine or methionine radioactivity in medium polypeptide. Schwann cells grown in a serum-free defined medium, in which schwann cells do not relate normally to axons, release approximately four times less labeled medium polypeptides tha cultures grown in medium supplemented with serum and chick embryo extract. In addition, there is a qualitative difference in the pattern of medium polypeptides resolved by SDS-PAGE, so that a single polypeptide (mol wt 40,000) accounts for nearly all of the label in medium polypeptides. Switching of cultures grown in defined medium to supplemented medium for 2 d results in a fourfold increase in the amount of labeled polypeptides appearing in the culture medium, and a return to the normal pattern of medium polypeptides appearing in the culture medium, and a return to the normal pattern of medium polypeptides as resolved by SDS-PAGE. This change in the pattern of polypeptides release by schwann cells is accompanied by changes in the association between schwann cells and axons. An early step in the establishment of normal axon-schwann cell relations appears to be an inward migration of schwann cells into axonal bundles and spreading of schwann cells along neurites. These changes are evident within 48 h after medium shift. Our results thus suggest that the release of proteins by schwann cells may be important for the development of normal axonal ensheathment.

scholarly journals Effect of FSH and cell localization on dimeric inhibin-A secretion from bovine granulosa cells in culture

2000 ◽  
Vol 165 (2) ◽  
pp. 207-215 ◽  
Author(s):  
ML Boudjemaa ◽  
P Rouillier ◽  
B Bhatia ◽  
JM Silva ◽  
LA Guilbault ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Culture System ◽  
Culture Medium ◽  
Cell Function ◽  
Spatial Differentiation ◽  
Defined Medium ◽  
Inhibin A ◽  
Mural Cells ◽  
Cell Localization ◽  
Molecular Masses

We tested the hypotheses that the secretion of dimeric inhibin-A from cultured bovine granulosa cells is stimulated by FSH, and that antral cells secrete more inhibin-A than do mural cells. Cells from the antral or mural compartment of follicles were cultured in defined medium in two culture systems, and dimeric inhibin-A was measured by two-site ELISA or by Western immunoblotting. In the first culture system, dimeric inhibin-A secretion declined with time in culture, but was significantly (P<0.05) higher from antral than from mural cells (as was total inhibin-alpha measured by RIA). The secretion of dimeric inhibin-A and inhibin-alpha from antral but not mural cells was responsive to FSH. In the second culture system, dimeric inhibin-A secretion increased with time in culture, and was significantly stimulated by FSH, but FSH responsiveness was dependent on the concentrations of insulin in the culture medium. The major forms of inhibin-A secreted had molecular masses of approximately 58, 62, 103-116 and >116 kDa; the 32 kDa form was barely detectable. These different forms were all stimulated by FSH, but the >116 and 62 kDa forms were most responsive to FSH. We conclude that (i) FSH stimulates dimeric inhibin-A secretion from bovine granulosa cells, (ii) the 62 kDa form of inhibin-A may be more responsive to FSH than the 58 kDa form, and (iii) the spatial differentiation of granulosa cell function within the follicle previously observed for oestradiol secretion was also observed for inhibin-alpha and dimeric inhibin-A secretion.

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