Selection of an optimal culture medium and the most responsive viability assay to assess AgNPs toxicity with primary cultures of Eisenia fetida coelomocytes

2019 ◽  
Vol 183 ◽  
pp. 109545 ◽  
Author(s):  
N. Garcia-Velasco ◽  
A. Irizar ◽  
E. Urionabarrenetxea ◽  
J.J. Scott-Fordsmand ◽  
M. Soto
Keyword(s):  
Eisenia Fetida ◽  
Culture Medium ◽  
Primary Cultures ◽  
Viability Assay ◽  
Selection Of

Pulsatile GnRH secretion from primary cultures of sheep olfactory placode explants

Reproduction ◽  
2000 ◽  
pp. 391-396 ◽  
Author(s):  
AH Duittoz ◽  
M Batailler
Keyword(s):  
Culture Medium ◽  
Primary Cultures ◽  
Pulsatile Secretion ◽  
Olfactory Placode ◽  
Gnrh Secretion ◽  
Individual Culture ◽  
Pulsatile Gnrh

The aim of this study was to investigate the development of pulsatile GnRH secretion by GnRH neurones in primary cultures of olfactory placodes from ovine embryos. Culture medium was collected every 10 min for 8 h to detect pulsatile secretion. In the first experiment, pulsatile secretion was studied in two different sets of cultures after 17 and 24 days in vitro. In the second experiment, a set of cultures was tested after 10, 17 and 24 days in vitro to investigate the development of pulsatile GnRH secretion in each individual culture. This study demonstrated that (i) primary cultures of GnRH neurones from olfactory explants secreted GnRH in a pulsatile manner and that the frequency and mean interpulse duration were similar to those reported in castrated ewes, and (ii) pulsatile secretion was not present at the beginning of the culture but was observed between 17 and 24 days in vitro, indicating the maturation of individual neurones and the development of their synchronization.

Abstract 261: Endothelial Cells Contribute to RAS Activation in Microvascular Smooth Muscle Cells

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Kayoko Miyata ◽  
Ryousuke Satou ◽  
L Gabriel Navar
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Mrna Expression ◽  
Culture Medium ◽  
Primary Cultures ◽  
Mrna Levels ◽  
Ang Ii ◽  
Relative Ratio ◽  
Renin Mrna ◽  
Afferent Arterioles

Introduction: We have demonstrated that Ang II augments angiotensinogen (AGT) expression in rat preglomerular vascular smooth muscle cells (VSMCs). However, it is unclear if endothelial cells (ECs) are involved in augmentation of AGT in renal afferent arterioles. Hypothesis: We assessed the hypothesis that the ECs respond to paracrine signals that Ang II contribute to AGT augmentation in VSMCs. Objective: We established primary cultures of preglomerular ECs and examined the effects of Ang II and/or culture medium from ECs on AGT expression in preglomerular VSMCs. Methods and Results: We established primary cultures of preglomerular ECs, isolated from afferent arterioles of Sprague-Dawley rats. The cells were identified as ECs by being positive for a marker, CD34 and endothelial NOS and negative for alpha-SMA (a marker for VSMCs) and P4H-b (a marker for Fibroblasts) by immnostaining. The expression levels of AGT mRNA and renin mRNA in preglomerular ECs were examined by real-time RT-PCR. Ang II (100 pmol/L) increased AGT mRNA levels (1.34 +/- 0.16, by 100 pmol/L, N=4) and Renin mRNA levels (6.16 +/- 0.96, by 100 nmol/L, N=4) in ECs. On the other hand, the same dose of Ang II suppressed Renin mRNA expression in isolated Juxtaglomerular cells (JGs). These results indicate that preglomerular ECs are respond to Ang II and exclude the possible contamination of JGs into ECs. 100 pmol/L of Ang II increased AGT mRNA expression levels (1.37 +/- 0.03, relative ratio, N=4) in preglomerular VSMCs and the culture medium of ECs without Ang II treatment also more increased AGT mRNA expression (1.62 +/- 0.13, relative ratio, N=4) in preglomerular VSMCs. The AGT mRNA expression augmentation was enhanced when preglomerular VSMCs were treated with culture medium of Ang II-treated preglomerular ECs (2.39 +/- 0.41, relative ratio, N=4). The synergistic effects of Ang II and preglomerular ECs were also observed in PAI-1 expression in preglomerular VSMCs. Conclusion: These data demonstrate that preglomerular ECs contribute to Ang II-upregulation of AGT in renal afferent arterioles leading to further Ang II augmentation, which leads to increases in inflammatory and sclerotic factors in preglomerular VSMCs.

Induction and regulation of connexin26 by glucagon in primary cultures of adult rat hepatocytes

1995 ◽  
Vol 108 (8) ◽  
pp. 2771-2780 ◽  
Author(s):  
T. Kojima ◽  
T. Mitaka ◽  
Y. Shibata ◽  
Y. Mochizuki
Keyword(s):  
Culture Medium ◽  
Primary Cultures ◽  
Minor Component ◽  
Rat Hepatocytes ◽  
Primary Hepatocytes ◽  
Adult Rat ◽  
Adult Rat Hepatocytes ◽  
Epidermal Growth ◽  
A Minor ◽  
Junction Proteins

In the adult rat hepatocyte, the gap junction proteins consist of a major component, connexin32 (Cx32) and a minor component, connexin26 (Cx26). Although we recently reported our success in inducing and maintaining Cx32 in adult rat hepatocytes cultured in serum-free L-15 medium supplemented with epidermal growth factor and 2% dimethyl sulfoxide, it was very difficult to induce Cx26 in the primary hepatocytes. In the present study, we found that the addition of 10(−7) M glucagon into the culture medium could dramatically induce Cx26 mRNA and protein. Although the expression of Cx32 mRNA was also influenced by glucagon, the increase of the expression was small. Immunocytochemically, Cx26-positive spots were observed between most adjacent cells and were co-localized with the Cx32-positive spots. We also examined whether 0.5 mM dibutyl cyclic AMP could induce expression of Cx26 in the cells. The effect of dexamethasone on the expression of Cx26 mRNA compared to that of Cx32 mRNA was examined. For the induction and maintenance of Cx26 mRNA, more than 10(−7) M dexamethasone was necessary in this culture. These results suggest that expression of Cx26 in hepatocytes may be regulated by the concentrations of glucagon and glucocorticoid hormones.

Proliferation and agglutinability of primary and transformed human epithelial cells in culture

1976 ◽  
Vol 21 (3) ◽  
pp. 553-561
Author(s):  
M.A. Ricard ◽  
R.J. Hay
Keyword(s):  
Culture Medium ◽  
Monolayer Culture ◽  
Primary Cultures ◽  
Short Term ◽  
Chloromethyl Ketone ◽  
Normal Populations ◽  
Short Term Exposure ◽  
Cell Strains ◽  
Human Placentae

Primary epithelial populations (HAM) were obtained by dissociation of the amniotic membrane stripped from human placentae. Agglutinability of cells from such normal populations and of cells from the transformed epithelial line WISH was then compared using concavanalin A as mediator. Extensive similar studies have previously been reported with cell strains isolated from other species. Freshly dissociated HAM cells from primary cultures agglutinated much less readily than did cells from WISH populations. Furthermore, the former exhibited a drastic decline in agglutinability as a function of time in suspension culture after trypsinization. Short-term exposure (60 h) of HAM cells in monolayer culture to 5-bromodeoxyuridine (BrdU) elicited heightened agglutinability detectable through 22 days in vitro. Addition of the protease inhibitors n-tosyl-L-lysyl-chloromethyl ketone (TLCK) or p-tosyl-L-arginine-methyl ester (TAME) to the culture medium inhibited proliferation of the WISH line by 40–50% while effecting only a 10–15% inhibition of HAM cells. These results also confirm data with other cell species indicating that high proteolytic activity at the surface of transformed cells may be related to the rapid proliferation rate.

scholarly journals Selection of Trichoderma spp. strains for the control of Sclerotinia sclerotiorum in soybean

2017 ◽  
Vol 52 (12) ◽  
pp. 1140-1148 ◽  
Author(s):  
Patrícia Elias Haddad ◽  
Luis Garrigós Leite ◽  
Cleusa Maria Mantovanello Lucon ◽  
Ricardo Harakava
Keyword(s):  
Sclerotinia Sclerotiorum ◽  
Culture Medium ◽  
Harmful Effect ◽  
Soybean Seeds ◽  
Trichoderma Spp ◽  
Negative Effect ◽  
Soybean Plants ◽  
Selection Of

Abstract: The objective of this work was to evaluate, in vitro and in vivo, the potential of Trichoderma spp. strains to control Sclerotinia sclerotiorum in soybeans (Glycine max) and to perform the molecular identification of the best perfoming strains. The effect of 120 strains of Trichoderma spp. on the viability of S. sclerotiorum sclerotia was evaluated in vitro through immersion in suspension of conidia from the antagonists and plating in culture medium. The best performing strains were evaluated in vivo, in a greenhouse, for control of the pathogen inoculated on 'Pintado' soybean seeds and plants. Of the 120 strains tested in vitro, 22 strains of Trichoderma spp. caused 100% inhibition of sclerotia germination. In the greenhouse, five strains inhibited the negative effect of the pathogen on seed germination and two strains increased in up to 67% plant dry matter. The best performing strains were identified as T. koningiopsis (3 strains), T. asperelloides (3), T. atroviride (2), and T. virens (1). Trichoderma strains are able to protect soybean plants from the harmful effect of S. sclerotiorum and, at the same time, they can promote the growth of the aerial part in greenhouse conditions.

scholarly journals Selection of filamentous fungi that are resistant to the herbicides atrazine, glyphosate and pendimethalin

2021 ◽  
Vol 43 ◽  
pp. e51656
Author(s):  
Nara Priscila Barbosa Bravim ◽  
Anatércia Ferreira Alves ◽  
José Fábio França Orlanda ◽  
Patricia Barbosa Rodrigues Silva
Keyword(s):  
Filamentous Fungi ◽  
Mycelial Growth ◽  
Culture Medium ◽  
Agricultural Soils ◽  
Culture Media ◽  
Fusarium Verticillioides ◽  
Fungal Growth ◽  
Relative Growth ◽  
Maximum Inhibition ◽  
Selection Of

The objective of the present study was to isolate fungi from agricultural soils and evaluate fungal growth in culture medium contaminated with atrazine, glyphosate and pendimethalin. Filamentous fungi were isolated from agricultural soils and cultured in a modified culture medium containing 0, 10, 20, 50, and 100 μg mL-1 atrazine, glyphosate and pendimethalin for 14 days at 28°C. The fungi that presented optimal and satisfactory growth were plated in Sabouraud culture medium with 4% dextrose and containing the herbicides at concentrations of 0, 10, 20, 50, and 100 μg mL-1 for seven days at 28°C. The mean mycelial growth values were submitted to analysis of variance and the Tukey test (p < 0.05%) for comparison and relative growth determination, and maximum inhibition rates were calculated. The isolated fungi Aspergillus fumigatus, Fusarium verticillioides and Penicillium citrinum were shown to be resistant to atrazine, glyphosate and pendimethalin. F. verticillioides showed higher mean mycelial growth in the culture media contaminated with atrazine and glyphosate than the other two fungi. In the culture medium contaminated with pendimethalin, F. verticillioides, and A. fumigatus presented the highest mean mycelial growth values.

scholarly journals Screening of Amazonian bacillus strains for lipase production using glycerol as substrate

ScientiaTec ◽  
2020 ◽  
Vol 7 (03) ◽  
Author(s):  
Giandra Volpato ◽  
Victória Furtado Migliavacca ◽  
Bruna Coelho de Andrade ◽  
Júlio Xandro Heck ◽  
Marco Antônio Záchia Ayub
Keyword(s):  
Organic Synthesis ◽  
Culture Medium ◽  
Lipolytic Activity ◽  
Gum Arabic ◽  
Lipase Production ◽  
Culture Conditions ◽  
Search And Selection ◽  
Triton X ◽  
Potential Use ◽  
Selection Of

The industrial application of lipolytic enzymes has been studied mainly due to the ability of these enzymes in catalyze reactions of synthesis and their stability in various organic solvents. One possibility is the use of lipase the organic synthesis, taking advantage as the generation of waste and difficult recovery of sub bioproducts. In this work, we carried out a selection of eighty-four isolates of Bacillus amazonian for lipase production, of which 30 strains showed lipolytic activity. The study of the culture conditions was performed through a Plackett-Burman experimental design using the strain that presented the highest lipolytic activity in a culture medium using glycerol as substrate.  The studied conditions were: concentration of soybean oil, olive oil, triton X-100, gum arabic, glycerol, and (NH4)2SO4, pH, temperature and concentration of inoculums. The best result obtained were 27 U/L in 48 h of cultivation by Bacillus circulans BL53. This work shows that the search and selection of microorganism with lipolytic activities can facilitate the discovery of new lipases, with potential use as by-product surplus.

scholarly journals Selection of the optimal culture medium for cultivation Fusarium sporotrichioides Sherb

2020 ◽  
Author(s):  
I.L. Astapchuk ◽  
◽  
N.A. Marchenko ◽  
G.V. Yakuba ◽  
A.I. Nasonov ◽  
...  
Keyword(s):  
Growth Rate ◽  
Growth And Development ◽  
Maximum Degree ◽  
Culture Medium ◽  
Culture Media ◽  
High Variability ◽  
Fusarium Sporotrichioides ◽  
Cultural Characteristics ◽  
Colony Diameter ◽  
Selection Of

The influence of various culture media on the growth, morphological and cultural characteristics of the fungus F. sporotrichioides was studied. Ten culture media were used in our research. A comparative study of the growth rate of the F. sporotrichioides mycelium made it possible to identify two media that are the most suitable for the cultivation and identification of this species, namely carrot and tomato agar. We took into account such criteria as ensuring the maximum degree of sporulation, rapid growth and development of mycelium (the 7th day), colony diameter (71–78 mm), as well as the ease of preparation. Nirenberg culture medium can be used to obtain a large number of conidia of the fungus. Because of the high variability of cultural characteristics of F. sporotrichioides, we recommend using different composition of media.

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