scholarly journals Achieving Shaker Flask-Scale Cell Densities in Miniprep-Scale Cultures with Baffled Culture Tubes and Growth Medium H15

BioTechniques ◽  
2000 ◽  
Vol 29 (6) ◽  
pp. 1207-1209
Author(s):  
L.J. Vigil ◽  
P.B. Danielson ◽  
C. Sollars ◽  
T.M. Yee ◽  
J.C. Fogleman
Keyword(s):  
Growth Medium ◽  
Shaker Flask ◽  
Cell Densities

scholarly journals Phenazine-1-Carboxamide Production in the Biocontrol Strain Pseudomonas chlororaphis PCL1391 Is Regulated by Multiple Factors Secreted into the Growth Medium

2001 ◽  
Vol 14 (8) ◽  
pp. 969-979 ◽  
Author(s):  
Thomas F. C. Chin-A-Woeng ◽  
Daan van den Broek ◽  
Gert de Voer ◽  
Koen M. G. M. van der Drift ◽  
Sietske Tuinman ◽  
...  
Keyword(s):  
Growth Medium ◽  
Homoserine Lactone ◽  
Dependent Manner ◽  
Pseudomonas Chlororaphis ◽  
In Vitro Production ◽  
Expression Studies ◽  
Multiple Copies ◽  
Cell Densities ◽  
Vitro Production

Pseudomonas chlororaphis PCL1391 controls tomato foot and root rot caused by Fusarium oxysporum f. sp. radicislycopersici. The production of phenazine-1-carboxamide (PCN) is crucial for this biocontrol activity. In vitro production of PCN is observed only at high-population densities, suggesting that production is under the regulation of quorum sensing. The main autoinducer molecule produced by PCL1391 was identified structurally as N-hexanoyl-l-homoserine lactone (C6-HSL). The two other autoinducers that were produced comigrate with N-butanoyl-l-homoserine lactone (C4-HSL) and N-octanoyl-l-homoserine lactone (C8-HSL). Two PCL1391 mutants lacking production of PCN were defective in the genes phzI and phzR, respectively, the nucleotide sequences of which were determined completely. Production of PCN by the phzI mutant could be complemented by the addition of exogenous synthetic C6-HSL, but not by C4-HSL, C8-HSL, or any other HSL tested. Expression analyses of Tn5luxAB reporter strains of phzI, phzR, and the phz biosynthetic operon clearly showed that phzI expression and PCN production is regulated by C6-HSL in a population density-dependent manner. The introduction of multiple copies of the regulatory genes phzI and phzR on various plasmids resulted in an increase of the production of HSLs, expression of the PCN biosynthetic operon, and consequently, PCN production, up to a sixfold increase in a copy-dependent manner. Surprisingly, our expression studies show that an additional, yet unidentified factor(s), which are neither PCN nor C4-HSL or C8-HSL, secreted into the growth medium of the overnight cultures, is involved in the positive regulation of phzI, and is able to induce PCN biosynthesis at low cell densities in a growing culture, resulting in an increase of PCN production.

Ultrastructural Alterations in Hela Cells Induced by Spider Venom

1967 ◽  
Vol 25 ◽  
pp. 20-21
Author(s):  
Dale E. McClendon ◽  
Paul N. Morgan ◽  
Bernard L. Soloff
Keyword(s):  
Growth Medium ◽  
Mammalian Cells ◽  
Carcinoma Cells ◽  
Venom Protein ◽  
Sterile Saline ◽  
Carbon Coated ◽  
Available Information ◽  
Loxosceles Reclusa ◽  
Essential Growth ◽  
Solution Cultures

It has been observed that minute amounts of venom from the brown recluse spider, Loxosceles reclusa, are capable of producing cytotoxic changes in cultures of certain mammalian cells (Morgan and Felton, 1965). Since there is little available information concerning the effect of venoms on susceptible cells, we have attempted to characterize, at the electron microscope level, the cytotoxic changes produced by the venom of this spider.Cultures of human epithelial carcinoma cells, strain HeLa, were initiated on sterile, carbon coated coverslips contained in Leighton tubes. Each culture was seeded with approximately 1x105 cells contained in 1.5 ml of a modified Eagle's minimum essential growth medium prepared in Hank's balanced salt solution. Cultures were incubated at 36° C. for three days prior to the addition of venom. The venom was collected from female brown recluse spiders and diluted in sterile saline. Protein determinations on the venom-were made according to the spectrophotometric method of Waddell (1956). Approximately 10 μg venom protein per ml of fresh medium was added to each culture after discarding the old growth medium. Control cultures were treated similarly, except that no venom was added. All cultures were reincubated at 36° C.

Effect of Iron Limitation on the Ultrastructure of Agmenellum Quadruplicatum

1981 ◽  
Vol 39 ◽  
pp. 410-411
Author(s):  
L. P. Hardie ◽  
D. L. Balkwill ◽  
S. E. Stevens
Keyword(s):  
Growth Medium ◽  
Green Alga ◽  
Natural Habitat ◽  
Iron Limitation ◽  
Natural Environments ◽  
Blue Green Alga ◽  
Marine Cyanobacterium ◽  
Natural Systems ◽  
Thin Sectioning ◽  
Agmenellum Quadruplicatum

Agmenellum quadruplicatum is a unicellular, non-nitrogen-fixing, marine cyanobacterium (blue-green alga). The ultrastructure of this organism, when grown in the laboratory with all necessary nutrients, has been characterized thoroughly. In contrast, little is known of its ultrastructure in the specific nutrient-limiting conditions typical of its natural habitat. Iron is one of the nutrients likely to limit this organism in such natural environments. It is also of great importance metabolically, being required for both photosynthesis and assimilation of nitrate. The purpose of this study was to assess the effects (if any) of iron limitation on the ultrastructure of A. quadruplicatum. It was part of a broader endeavor to elucidate the ultrastructure of cyanobacteria in natural systemsActively growing cells were placed in a growth medium containing 1% of its usual iron. The cultures were then sampled periodically for 10 days and prepared for thin sectioning TEM to assess the effects of iron limitation.

Automated cell counting of astrocytes on patterned substrates containing aliphatic and charged properties

1995 ◽  
Vol 53 ◽  
pp. 974-975
Author(s):  
W. Shain ◽  
H. Ancin ◽  
H.C. Craighead ◽  
M. Isaacson ◽  
L. Kam ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Attachment ◽  
Culture Method ◽  
Cell Counting ◽  
Data Sets ◽  
Nuclear Staining ◽  
Double Positive ◽  
A Cell ◽  
Wafer Test ◽  
Cell Densities

Neural protheses have potential to restore nervous system functions lost by trauma or disease. Nanofabrication extends this approach to implants for stimulating and recording from single or small groups of neurons in the spinal cord and brain; however, tissue compatibility is a major limitation to their practical application. We are using a cell culture method for quantitatively measuring cell attachment to surfaces designed for nanofabricated neural prostheses.Silicon wafer test surfaces composed of 50-μm bars separated by aliphatic regions were fabricated using methods similar to a procedure described by Kleinfeld et al. Test surfaces contained either a single or double positive charge/residue. Cyanine dyes (diIC18(3)) stained the background and cell membranes (Fig 1); however, identification of individual cells at higher densities was difficult (Fig 2). Nuclear staining with acriflavine allowed discrimination of individual cells and permitted automated counting of nuclei using 3-D data sets from the confocal microscope (Fig 3). For cell attachment assays, LRM5 5 astroglial cells and astrocytes in primary cell culture were plated at increasing cell densities on test substrates, incubated for 24 hr, fixed, stained, mounted on coverslips, and imaged with a 10x objective.

Effects of resuspension of eastern oyster Crassostrea virginica biodeposits on phytoplankton community structure

2020 ◽  
Vol 640 ◽  
pp. 79-105
Author(s):  
ET Porter ◽  
E Robins ◽  
S Davis ◽  
R Lacouture ◽  
JC Cornwell
Keyword(s):  
Water Quality ◽  
Community Structure ◽  
Crassostrea Virginica ◽  
Skeletonema Costatum ◽  
Eastern Oyster ◽  
Energy Dissipation Rate ◽  
Negative Effects ◽  
Cell Densities ◽  
Sediment Bottom ◽  
Nitrite Nitrate

Anthropogenic disturbances in the Chesapeake Bay (USA) have depleted eastern oyster Crassostrea virginica abundance and altered the estuary’s environment and water quality. Efforts to rehabilitate oyster populations are underway; however, the effect of oyster biodeposits on water quality and plankton community structure are not clear. In July 2017, we used 6 shear turbulence resuspension mesocosms (STURMs) to determine differences in plankton composition with and without the daily addition of oyster biodeposits to a muddy sediment bottom. STURM systems had a volume-weighted root mean square turbulent velocity of 1.08 cm s-1, energy dissipation rate of ~0.08 cm2 s-3, and bottom shear stress of ~0.36-0.51 Pa during mixing-on periods during 4 wk of tidal resuspension. Phytoplankton increased their chlorophyll a content in their cells in response to low light in tanks with biodeposits. The diatom Skeletonema costatum bloomed and had significantly longer chains in tanks without biodeposits. These tanks also had significantly lower concentrations of total suspended solids, zooplankton carbon, and nitrite +nitrate, and higher phytoplankton carbon concentrations. Results suggest that the absence of biodeposit resuspension initiates nitrogen uptake for diatom reproduction, increasing the cell densities of S. costatum. The low abundance of the zooplankton population in non-biodeposit tanks suggests an inability of zooplankton to graze on S. costatum and negative effects of S. costatum on zooplankton. A high abundance of the copepod Acartia tonsa in biodeposit tanks may have reduced S. costatum chain length. Oyster biodeposit addition and resuspension efficiently transferred phytoplankton carbon to zooplankton carbon, thus supporting the food web in the estuary.

Use of multicriterial optimization of the growth medium for accumulation of biomass of lactic acid bacteria

2020 ◽  
Vol 26 (4) ◽  
pp. 47-57
Author(s):  
М. Khonkiv ◽  
S. Teterina ◽  
S. Danylenko ◽  
O. Potemska
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Growth Medium ◽  
Multicriterial Optimization

Cultivation and Harvesting of Selenium-Enriched Ganoderma lingzhi and Spent Medium Using Kudzu Vine as Substrate

2019 ◽  
Vol 18 (2) ◽  
pp. 152-157
Author(s):  
Zeng Xianlu ◽  
Han Fei ◽  
Zhong Yanmei
Keyword(s):  
Growth Medium ◽  
Wheat Bran ◽  
Fruiting Body ◽  
Sodium Selenite ◽  
Human Consumption ◽  
Growth Substrate ◽  
Organic Selenium ◽  
Significant Difference ◽  
Ganoderma Lingzhi ◽  
Run Speed

In order to harvest selenium-enriched fruiting body and spores of Ganoderma lingzhi and spent medium, G. lingzhi was cultivated in kudzu vine as substrate and the bio-transformation of selenite was evaluated. The growth medium consisted of Kudzu vine supplemented with 20% wheat bran or sawdust or none. The growth medium was supplemented with 0, 10, 20, 30, and 50 mg/kg of sodium selenite. We found a significant difference in spawn run speed, fruiting body and spore yields when Kudzu vine was supplemented with wheat bran or sawdust. However, when whole-kudzu vine was used alone as substrate, it resulted in a significantly lower spawn run speed, fruiting body, and spore yields compared with kudzu vine + sawdust substrate and kudzu vine + wheat bran substrate. The selenium content in fruiting body and spores increased with increasing sodium selenite supplementation and approximately equaled half of the selenium in the substrate. No selenite was detected in both the fruiting body and spores. However, in the spent medium when sodium selenite was supplemented at 10, 20, 30, 50 mg/kg, the residual selenite concentration decreased to 0.45, 0.72, 1.29, and 1.95 mg/kg, respectively, suggesting a higher selenite transformation (92.27–93.57%). In conclusion, if Ganoderma fruiting body and spores were to be harvested for human consumption, approximately 50 mg/kg selenite should be added to the growth substrate. On the other hand, if the spent medium was to be used as an organic selenium source, the optimal sodium selenite supplementation level would be 10 mg/kg.

Selenium Uptake by a Coal Mine Wetland Sediment

2003 ◽  
Vol 38 (3) ◽  
pp. 483-497 ◽  
Author(s):  
Susan A. Baldwin ◽  
Al Henry Hodaly
Keyword(s):  
Growth Medium ◽  
Coal Mine ◽  
Mine Waste ◽  
Reduction Rate ◽  
Plant Debris ◽  
Bacteria Growth ◽  
Sulfate Reducing ◽  
Selenium Uptake ◽  
Negative Effect ◽  
Reducing Bacteria

Abstract Sediment from a wetland receiving runoff from a coal mine waste dump in the Elk River Valley of southeast British Columbia was assessed for potential selenium uptake. Selenite [SeO32-, Se(IV)] was found to adsorb to the washed sediment at pH 7 to 8, whereas no selenate [SeO42-, Se(VI)] was adsorbed, in the concentration range of 8 to 225 μg L-1 Se as selenite or selenate. Sulfate- and selenate-reducing bacterial activity was detected in the sediment. In the presence of sulfate-reducing bacteria growth medium, Se as selenate was reduced from 619(±53) μg L-1 to 15(±0.7) μg L-1, and in the presence of selenate-reducing bacteria growth medium, Se as selenate was reduced from 364(±66) mg L-1 to 22(±10) mg L-1. Semi-continuous microcosms containing sediment overlaid with selenate (500 μg L-1 Se) and sulfate (0.9 g L-1) containing water were amended with plant debris from the site or nutrients (lactate and fertilizer). Potential selenate reduction rate (0.76 h-1) was highest in the unamended microcosms. Amendment with plant debris from the site had a negative effect on selenate reduction rate in the short term (after one hour) and a positive effect on Se removal in the long term (after one week). This study suggests that wetland sediments at the mine site may be important sinks for Se.

scholarly journals High-throughput microbioreactor provides a capable tool for early stage bioprocess development

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mathias Fink ◽  
Monika Cserjan-Puschmann ◽  
Daniela Reinisch ◽  
Gerald Striedner
Keyword(s):  
High Throughput ◽  
Process Development ◽  
Early Stage ◽  
Explanatory Power ◽  
Batch Mode ◽  
E Coli ◽  
Speed Up ◽  
Bioreactor System ◽  
Cell Densities ◽  
Microbial Systems

AbstractTremendous advancements in cell and protein engineering methodologies and bioinformatics have led to a vast increase in bacterial production clones and recombinant protein variants to be screened and evaluated. Consequently, an urgent need exists for efficient high-throughput (HTP) screening approaches to improve the efficiency in early process development as a basis to speed-up all subsequent steps in the course of process design and engineering. In this study, we selected the BioLector micro-bioreactor (µ-bioreactor) system as an HTP cultivation platform to screen E. coli expression clones producing representative protein candidates for biopharmaceutical applications. We evaluated the extent to which generated clones and condition screening results were transferable and comparable to results from fully controlled bioreactor systems operated in fed-batch mode at moderate or high cell densities. Direct comparison of 22 different production clones showed great transferability. We observed the same growth and expression characteristics, and identical clone rankings except one host-Fab-leader combination. This outcome demonstrates the explanatory power of HTP µ-bioreactor data and the suitability of this platform as a screening tool in upstream development of microbial systems. Fast, reliable, and transferable screening data significantly reduce experiments in fully controlled bioreactor systems and accelerate process development at lower cost.

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