scholarly journals Optimizing design and fabrication of microfluidic devices for cell cultures: An effective approach to control cell microenvironment in three dimensions

2014 ◽  
Vol 8 (4) ◽  
pp. 046503 ◽  
Author(s):  
G. Pagano ◽  
M. Ventre ◽  
M. Iannone ◽  
F. Greco ◽  
P. L. Maffettone ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Control Cell ◽  
Microfluidic Devices ◽  
Three Dimensions ◽  
Cell Microenvironment ◽  
Optimizing Design

scholarly journals Generating linear oxygen gradients across 3D cell cultures with block-layered oxygen controlled chips (BLOCCs)

2020 ◽  
Vol 12 (1) ◽  
pp. 18-24 ◽  
Author(s):  
Matthew W. Boyce ◽  
William C. Simke ◽  
Rachael M. Kenney ◽  
Matthew R. Lockett
Keyword(s):  
Cell Cultures ◽  
Cell Microenvironment ◽  
3D Cell Cultures ◽  
Oxygen Gradients

BLOCCs are readily assembled structures of laser cut acrylic and silicone, capable of imposing physiologically relevant oxygen gradients across 3D cell cultures. With sensors and cell-based readouts, we quantified cell-microenvironment relationships.

Fabrication of composite microfluidic devices for local control of oxygen tension in cell cultures

Lab on a Chip ◽  
2019 ◽  
Vol 19 (2) ◽  
pp. 306-315 ◽  
Author(s):  
Yandong Gao ◽  
Gulnaz Stybayeva ◽  
Alexander Revzin
Keyword(s):  
Oxygen Tension ◽  
Cell Cultures ◽  
Local Control ◽  
Microfluidic Chip ◽  
Gas Permeability ◽  
Microfluidic Devices

We developed a microfabrication strategy that integrated two materials with different gas permeability in a single microfluidic chip to enable local control of oxygen tension for cell cultures.

scholarly journals Genetically Encoded Red Photosensitizers with Enhanced Phototoxicity

2020 ◽  
Vol 21 (22) ◽  
pp. 8800 ◽  
Author(s):  
Dmitry A. Gorbachev ◽  
Dmitry B. Staroverov ◽  
Konstantin A. Lukyanov ◽  
Karen S. Sarkisyan
Keyword(s):  
Directed Evolution ◽  
Cell Fate ◽  
Mammalian Cell ◽  
Cell Cultures ◽  
Fluorescent Protein ◽  
Control Cell ◽  
Red Fluorescent Protein ◽  
Mammalian Cell Cultures

Genetically encoded photosensitizers are increasingly used as optogenetic tools to control cell fate or trigger intracellular processes. A monomeric red fluorescent protein called SuperNova has been recently developed, however, it demonstrates suboptimal characteristics in most phototoxicity-based applications. Here, we applied directed evolution to this protein and identified SuperNova2, a protein with S10R substitution that results in enhanced brightness, chromophore maturation and phototoxicity in bacterial and mammalian cell cultures.

Effects of erythropoietin on uridine metabolism in cell cultures of fetal calf liver

1984 ◽  
Vol 62 (2-3) ◽  
pp. 143-149 ◽  
Author(s):  
L. F. Congote
Keyword(s):  
Cell Cultures ◽  
High Performance ◽  
Specific Activity ◽  
Control Cell ◽  
Fold Increase ◽  
Reversed Phase ◽  
Uridine Incorporation ◽  
Soluble Cell ◽  
Cell Extracts ◽  
Chain Synthesis

The effect of sheep plasma erythropoietin preparations on the incorporation of [5-3H]uridine into erythroid cells has been studied using cells of fetal calf liver cultured in serum-free medium. The cells were incubated for 20 h with the hormone, followed by a 1-h incubation with [3H]uridine. Erythropoietin caused a 2.5-fold increase in the incorporation of uridine into cold trichloroacetic-acid-insoluble cell extracts and a 70% increase in the incorporation of uridine into the cold acid-soluble cell extracts. The phosphorylated metabolites of labeled uridine present in the cold acid-soluble fraction were analyzed by anion-exchange high performance liquid chromatography (HPLC). Erythropoietin increased the amounts of labeled UDP and UTP per cell. However, the specific activity of UTP and the labeled amounts of UDP-glucose in erythropoietin-treated cells were not significantly different from those in control cell cultures. After chromatography of the crude erythropoietin preparations on reversed-phase and gel-permeation HPLC, there was a perfect coincidence of the fractions stimulating uridine incorporation into acid-soluble and acid-insoluble cell extracts. The protein fractions from crude erythropoetin which stimulated uridine incorporation after purification by reversed-phased HPLC were also able to stimulate globin chain synthesis in fetal calf liver cells. These experiments suggest that the multiple effects on uridine metabolism described above are due to erythropoietin, rather than other proteins contaminating the crude hormone preparations.

scholarly journals Toxicity of organic and inorganic nickel in pancreatic cell cultures: Comparison to cadmium

2020 ◽  
Vol 70 (6) ◽  
pp. 344-359
Author(s):  
David Wallace ◽  
Aleksandra Buha-Đorđević ◽  
Alexander Benton
Keyword(s):  
Cell Cultures ◽  
Viable Cell ◽  
Pancreatic Cell ◽  
Nickel Toxicity ◽  
Cell Microenvironment ◽  
Viable Cells ◽  
Nickel Compounds ◽  
Nickel Exposure ◽  
Cancerous Cells ◽  
Group 1

Nickel compounds are Group 1 carcinogens and possibly cancer-causing in the pancreas. We examined the toxicity of nickel in both 2-D and 3-D pancreatic cell cultures, to determine the LD50 for organic and inorganic nickel in normal and cancerous cells. Assays with cadmium chloride were performed to be a comparison to potential nickel-induced toxicity. Cells were exposed to twelve concentrations of NiCl2 or Ni-(Ac)2 for 48h (2-D), or six concentrations for 48 hours (3-D). There was a significant (P=0.0016) difference between HPNE and AsPC-1 LD50 values after cadmium exposure, at 69.9 µM and 29.2 µM, respectively. Neither form of nickel exhibited toxicity in 2-D or 3-D cultures, but after 48h, changes in spheroid morphology were observed. The inability of Ni to reduce viable cell numbers suggests a toxic mechanism that differs from cadmium, also a Group 1 carcinogen. The cell microenvironment was not a factor in nickel toxicity with no changes in viable cells in either 2-D or 3-D cultures. These studies only examined cytotoxicity, and not genotoxicity, a potential mechanism of nickel carcinogenicity. Alterations in DNA function or the expression of apoptotic proteins/processes would take longer to manifest. Current work focuses on cellular changes following extended nickel exposure.

scholarly journals PDMS-Based Microfluidic Devices for Cell Culture

Inventions ◽  
2018 ◽  
Vol 3 (3) ◽  
pp. 65 ◽  
Author(s):  
Stefania Torino ◽  
Brunella Corrado ◽  
Mario Iodice ◽  
Giuseppe Coppola
Keyword(s):  
Cell Culture ◽  
Cell Cultures ◽  
Soft Lithography ◽  
Microfluidic Devices ◽  
Biological Research ◽  
Culture Chamber ◽  
Culture Methods ◽  
Critical Issues ◽  
Scaling Down ◽  
Cell Culture Methods

Microfluidic technology has affirmed itself as a powerful tool in medical and biological research by offering the possibility of managing biological samples in tiny channels and chambers. Among the different applications, the use of microfluidics for cell cultures has attracted much interest from scientists worldwide. Traditional cell culture methods need high quantities of samples and reagents that are strongly reduced in miniaturized systems. In addition, the microenvironment is better controlled by scaling down. In this paper, we provide an overview of the aspects related to the design of a novel microfluidic culture chamber, the fabrication approach based on polydimethylsiloxane (PDMS) soft-lithography, and the most critical issues in shrinking the size of the system.

scholarly journals Somatostatin receptor gene expression and inhibitory effects of octreotide on primary cultures of orbital fibroblasts from Graves' ophthalmopathy

2000 ◽  
Vol 25 (1) ◽  
pp. 63-71 ◽  
Author(s):  
D Pasquali ◽  
P Vassallo ◽  
D Esposito ◽  
G Bonavolonta ◽  
A Bellastella ◽  
...  
Keyword(s):  
Cell Growth ◽  
Cell Cultures ◽  
Specific Binding ◽  
Somatostatin Receptor ◽  
Primary Cultures ◽  
Control Cell ◽  
Gene Transcript ◽  
Camp Accumulation ◽  
Graves Ophthalmopathy ◽  
Class 1

To explore the mechanism underlying the effects of the somatostatin (SST) analogue octreotide in Graves' ophthalmopathy (GO), we investigated the expression of SST and of SST receptor (sst(1-5)) genes in primary cultures of fibroblasts established from retroorbital tissue of GO patients and of control subjects. We determined also SST specific binding sites by competitive binding of [(125)ITyr(11)]SST-14 and the effect of octreotide on cell growth, cAMP accumulation, Bcl-2 intracellular levels and apoptosis in GO fibroblast primary cultures. All primary cultures expressed the SST gene transcript and one or more ssts that have a high affinity for the two analogues (class 1 sst. The sst(2) transcript was found in nine, sst(3) in five and sst(5) in eight out of ten GO cell cultures. Sst(2) was detected in all six, and sst(3) in four out of the six control cell cultures. Sst(4) was absent from all samples, and sst(1) was found only in six out of the ten GO samples. SST-14 and octreotide inhibited the binding of [(125)I-Tyr(11)]SST-14 with a half-maximal inhibition of binding (IC(50)) of 0.80+/-0.37 and 33. 7+/- 33.1 nmol/l respectively in GO cell cultures, and with an IC(50) of 0.9 and 1.5 nmol/l in control cultures. Octreotide (10(-6) and 10(-7) M) significantly decreased (P<0.001) forskolin-induced but not basal cAMP accumulation; at both doses for 72 h it inhibited cell growth (20 and 55% respectively), and induced apoptosis (20 and 40%), and abolished Bcl-2 protein in cell lysates. In conclusion, SST and sst transcripts are expressed and functional in cultured retroorbital fibroblasts. The presence of class 1 sst in GO tissue and the inhibition exerted by octreotide on retroorbital cell growth and activity in vitro may account for the effects of SST analogue administration in vivo in GO.

scholarly journals Tailoring Cellular Function: The Contribution of the Nucleus in Mechanotransduction

2021 ◽  
Vol 8 ◽  
Author(s):  
Fabrizio A. Pennacchio ◽  
Paulina Nastały ◽  
Alessandro Poli ◽  
Paolo Maiuri
Keyword(s):  
Control Cell ◽  
Cellular Responses ◽  
Mechanical Stimuli ◽  
Mechanical State ◽  
Cellular Functions ◽  
Cell Microenvironment ◽  
Cytoskeleton Organization ◽  
Pathological Conditions ◽  
Complex Process ◽  
Health And Disease

Cells sense a variety of different mechanochemical stimuli and promptly react to such signals by reshaping their morphology and adapting their structural organization and tensional state. Cell reactions to mechanical stimuli arising from the local microenvironment, mechanotransduction, play a crucial role in many cellular functions in both physiological and pathological conditions. To decipher this complex process, several studies have been undertaken to develop engineered materials and devices as tools to properly control cell mechanical state and evaluate cellular responses. Recent reports highlight how the nucleus serves as an important mechanosensor organelle and governs cell mechanoresponse. In this review, we will introduce the basic mechanisms linking cytoskeleton organization to the nucleus and how this reacts to mechanical properties of the cell microenvironment. We will also discuss how perturbations of nucleus–cytoskeleton connections, affecting mechanotransduction, influence health and disease. Moreover, we will present some of the main technological tools used to characterize and perturb the nuclear mechanical state.

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