Investigation of the Influences of Micro Vibrational Stimuli and Hydrophilicity of a Scaffold on a Bovine Endothelial Cell Culture

2010 ◽  
Author(s):  
Ching-Wen Li ◽  
Gou-Jen Wang
Keyword(s):  
Cultured Cells ◽  
Adhesive Force ◽  
Effective Solution ◽  
Endothelial Cell Culture ◽  
Bovine Endothelial Cell ◽  
Sheer Stress ◽  
Applied Shear Stress ◽  
Bovine Endothelial Cells ◽  
A Cell ◽  
Micro Vibration

Both the hydrophilicity of the scaffold and applied sheer stress can influence the growth of cultured cells. In this study, the influences of applied shear stress and the hydrophilicity of the scaffold on the growth of bovine endothelial cells (BEC) were investigated. A piezoelectric micro vibrational stage was used to provide micro vibrational stimuli of different frequencies to generate various sheer stresses. 24-well, biodegradable lactide-co-glycolide (PLGA) scaffolds, and nanostructuresd PLGA scaffolds were used for the cell culturing. From the results of this study, it can be inferred that the vibration induced sheer stress can effectively enhance BEC growth as long as the corresponding sheer force is less than the adhesive force between a cell and the scaffold. It is also suggested that micro vibration stimulus may be a more cost and time effective solution than the nanostructured scaffold approach for the enhancement of BEC growth.

A New In Vitro Model for Predicting the Toxicity of Cosmetic Products

1992 ◽  
Vol 20 (1) ◽  
pp. 138-143
Author(s):  
Maria Carrara ◽  
Lorenzo Cima ◽  
Roberto Cerini ◽  
Maurizio Dalle Carbonare
Keyword(s):  
Cell Culture ◽  
Cultured Cells ◽  
Neutral Red ◽  
Total Protein Content ◽  
Cosmetic Products ◽  
Cell Culture Insert ◽  
A Cell ◽  
Vitro Model ◽  
Cosmetic Emulsions

A method has been developed whereby cosmetic products which are not soluble in water or in alcohol can be brought into contact with cell cultures by being placed in a cell culture insert, which is then placed in the cell culture well. Preliminary experiments were carried out with L929 cells, and cytotoxicity was evaluated by measuring neutral red uptake and the total protein content of treated cultured cells. Encouraging results were obtained in comparisons of three cosmetic emulsions and of one emulsion containing a range of concentrations of two preservatives, Kathon CG and Bronopol.

INTERACTION BETWEEN CULTURED ENDOTHELIAL CELLS AND ABNORMAL ANTITHROMBIN III "TOYAMA"

1987 ◽  
Author(s):  
N Sakuragawa ◽  
S Saitoh ◽  
K Takahashi
Keyword(s):  
Endothelial Cells ◽  
Room Temperature ◽  
Antithrombin Iii ◽  
Cultured Cells ◽  
Binding Activity ◽  
Endothelial Cell Culture ◽  
Antithrombin Activity ◽  
Thrombin Activity ◽  
Cultured Endothelial Cells ◽  
At Iii

Purpose: Abnormal antithrombin III(AT-III)Toyama showed non-affinity to heparin and heparinoid to show loss of immediate antithrombin activity. On the endothelial cells, there are heparinoids including heparan sulfate. We investigated on the interaction between cultured endothelial cells and abnormal AT-III"Toyama" from the viewpoint of antithrombin activity.Materials and methods: (1) Endothelial cell culture:^125I-labelled normal and abnormal AT-III were placed on the washed endothelial cultured cells in 0.2 ml of RPMI-1640 medium for 15 min at 37°C. The medium was suctioned off and the cell layer was washed with Hank's balanced salt solution. The cells were incubated with 1 ml of heparin(3 ug/ml) for 15 min at 4°C. The radioactivity in the supernatant was counted, and represented AT-III which bound to the cells surface. (2) Antithrombin activity: 0.23 ml of thrombin solution^ U/ml) and 0.03 ml of normal or abnormal AT-III plasma were mixed, and incubated on the cultured cell surface for 5 min at room temperature. The residual thrombin activity was assayed by 0.3 ml of the substrate (S-2238) solution(0.8mM)for 5 min. After these procedures,2 ml of 2% citric acid solution was added to stop the reaction, and 0D(405 nm) was recorded.Results: Abnormal AT-III showed reduced binding-activity to cultured cells to one fifth compared with normal AT-III, and the residual thrombin activity in the abnormal was higher compared with that in normal plasma.Conclusion: Abnormal AT-III showed less binding activity to the cultured endothelial cells, and less thrombin neutralizing activity to show thrombogenic tendency.

Characterization of phospholipase D in a cell-free system of cultured cells derived from rat frontal cortex

1992 ◽  
Vol 595 (1) ◽  
pp. 12-16 ◽  
Author(s):  
Akira Nishida ◽  
Masami Shimizu ◽  
Yasunori Kanaho ◽  
Yoshinori Nozawa ◽  
Shigeto Yamawaki
Keyword(s):  
Frontal Cortex ◽  
Phospholipase D ◽  
Cultured Cells ◽  
Free System ◽  
Cell Free System ◽  
A Cell ◽  
Rat Frontal Cortex

Transfer-Print of CNTs and its Application to Cell Scaffold

2017 ◽  
Vol 11 (6) ◽  
pp. 941-946 ◽  
Author(s):  
Arata Kaneko ◽  
◽  
Yuuki Miyazaki ◽  
Tatsuya Goto
Keyword(s):  
Cultured Cells ◽  
Hydrophobic Interactions ◽  
Casting Process ◽  
Water Contact ◽  
Si Substrates ◽  
Dimethyl Siloxane ◽  
Cell Scaffold ◽  
Assembled Monolayers ◽  
A Cell ◽  
Surface Modified

A bio-chip using cultured cells is developed for an application to drug screening. Carbon nanotubes (CNTs) are a candidate for this electrode material. A transfer-prints is expected to be a CNT-patterning technique applicable to soft material. This present paper is intended to show some basic properties about the transfer-print of CNTs, and also to demonstrate the possibility of the CNTs as a cell scaffold. The present study prepared several types of surface-modified Si substrate with different wettability to investigate the effects of wettability on the transferring ratio of CNTs. Some Si substrates are terminated by OH or H groups, while other substrates are coated with hydrophobic or hydrophilic self-assembled monolayers. The stamps for transfer-print, which have circular dots (50-μm diameter) or a straight ridge (50-μm width) array, are fabricated using poly-dimethyl-siloxane (PDMS). The surfaces of PDMS stamps are inked by single-walled CNTs by a pre-transferring or casting process. The transfer-prints to surface-modified Si surfaces allow the CNTs to be formed in lines of several tens of micrometers, while the coverage of transfer-printed CNTs is also dominated by surface wettability. The coverage of transfer-printed CNTs increases with the water contact angle of the Si surface. It is reasonable that the transfer-print of CNTs is performed by hydrophobic interactions. Meanwhile, two kinds of polymer (polystyrene (PS) and polyethylene terephthalate (PET)) sheets are also utilized as a substrate. The transfer-prints with heating around the softening point of the polymer allow CNTs to be accurately patterned into an array of 50-μm dots. The coverage of CNTs is 94% on the PET substrate. The PS sheet with patterned CNTs is applied to a cell scaffold. PC12 cells are cultured on the PS sheets so that the cells are selectively adhered to the transfer-printed CNTs. The adhered cells are extended with some pseudopods. It is demonstrated that the transfer-printed CNTs are expected to be electrodes of the cell scaffold.

S-phase-specific expression of the Mad3 gene in proliferating and differentiating cells

2001 ◽  
Vol 359 (2) ◽  
pp. 361-367 ◽  
Author(s):  
Elizabeth J. FOX ◽  
Stephanie C. WRIGHT
Keyword(s):  
Cell Cycle ◽  
Cellular Proliferation ◽  
Cultured Cells ◽  
S Phase ◽  
Proliferating Cells ◽  
Specific Expression ◽  
Related Function ◽  
Erythroleukemia Cells ◽  
A Cell ◽  
Pass Through

The Myc/Max/Mad transcription factor network plays a central role in the control of cellular proliferation, differentiation and apoptosis. In order to elucidate the biological function of Mad3, we have analysed the precise temporal patterns of Mad3 mRNA expression during the cell cycle and differentiation in cultured cells. We show that Mad3 is induced at the G1/S transition in proliferating cells; expression persists throughout S-phase, and then declines as cells pass through G2 and mitosis. The expression pattern of Mad3 is coincident with that of Cdc2 throughout the cell cycle. In contrast, the expression of Mad3 during differentiation of cultured mouse erythroleukemia cells shows two transient peaks of induction. The first of these occurs at the onset of differentiation, and does not correlate with the S-phase of the cell cycle, whereas the second is coincident with the S-phase burst that precedes the terminal stages of differentiation. Our results therefore suggest that Mad3 serves a cell-cycle-related function in both proliferating and differentiating cells, and that it may also have a distinct role at various stages of differentiation.

scholarly journals Improving Membrane Staining of Cultured Cells Using Ferrocyanide as a Post-Fixative

2007 ◽  
Vol 15 (2) ◽  
pp. 38-39
Author(s):  
Stéphane Nizet
Keyword(s):  
Ultrathin Section ◽  
Cultured Cells ◽  
Cell Model ◽  
Cell Monolayer ◽  
Cancer Cell Line ◽  
Colon Cancer Cell Line ◽  
Colon Cancer Cell ◽  
Human Intestine ◽  
A Cell ◽  
Permeability Studies

Lack of contrast is a common problem encountered when doing TEM of cultured cells, especially of membranes. Using ferrocyanide as a post-fixative can greatly improve membrane fixation and staining. This protocol has been used to study Caco-2 cells grown on PET membranes (the “Caco model”). Caco-2 is a colon cancer cell line that differentiates upon reaching confluency. This allows permeability studies on a cell model, which is reasonably similar to the human intestine.Basically, the protocol is classical, the only peculiarity consisting in including ferrocyanide in post-fixation. I describe how I prepare and embed the membrane in order to obtain transverse sections of a cell monolayer because I find this is the only way to obtain regular sections with the cells sticking to the membrane (otherwise the ultrathin section splits between the cell and membrane).

scholarly journals Analysis of Mycobacterium Species for the Presence of a Macrolide Toxin, Mycolactone

2004 ◽  
Vol 72 (1) ◽  
pp. 123-132 ◽  
Author(s):  
Alexa K. Daniel ◽  
Richard E. Lee ◽  
Francoise Portaels ◽  
P. L. C. Small
Keyword(s):  
Cultured Cells ◽  
Cell Envelope ◽  
Buruli Ulcer ◽  
Fibroblast Cell ◽  
Mycobacterium Ulcerans ◽  
Mycobacterial Species ◽  
Murine Fibroblast ◽  
A Cell ◽  
Layer Chromatography ◽  
Slow Growing

ABSTRACT Mycobacterium ulcerans is an environmental organism which is responsible for the disease Buruli ulcer, a necrotizing skin disease emerging in west Africa. M. ulcerans produces the polyketide-derived macrolide mycolactone, which is required for the immunosuppression and tissue damage which characterizes Buruli ulcer. We have extracted lipids from the cell envelope and culture filtrate from 52 isolates of Mycobacterium species, analyzed them with thin-layer chromatography, and tested them in a murine fibroblast cell line (L929) cytotoxicity assay to investigate whether these mycobacterial species produce mycolactone. For these studies chloroform-methanol (2:1, vol/vol) extracts were prepared from representative fast- and slow-growing mycobacterial species. Isolates tested included 16 uncharacterized, slow-growing, environmental mycobacterial species isolated from areas in which M. ulcerans infection is endemic. Although several strains of mycobacteria studied produced cytopathic lipids, none of these produced a phenotype on cultured cells consistent with that produced by mycolactone. Two mycobacterial species, M. scrofulaceum and M. kansasii, and eight of the environmental mycobacterial isolates contained cell-associated lipids cytopathic to fibroblasts at concentrations of 33 to 1,000 μg/ml. In contrast, mycolactone produces cytotoxicity at less than 2 ng/ml. Analysis of 16S rRNA sequences from the eight environmental isolates suggests that these are novel mycobacterial species. Results from these studies suggest that, although production of cytopathic lipids is relatively common among mycobacterial species, the production of mycolactone as a cell-associated or secreted molecule appears so far to be restricted to M. ulcerans.

scholarly journals Characterization and simulation of peristaltic micropumps

2013 ◽  
Vol 2 (2) ◽  
pp. 165-169 ◽  
Author(s):  
M. Busek ◽  
M. Nötzel ◽  
C. Polk ◽  
F. Sonntag
Keyword(s):  
Cell Culture ◽  
Culture System ◽  
Cultured Cells ◽  
Applied Pressure ◽  
Cell Culture System ◽  
Heart Activity ◽  
Non Invasive ◽  
Micro Piv ◽  
Piv Method ◽  
A Cell

Abstract. The aim of the work is to find an analytical model of a pneumatic micropump which was integrated into a cell-culture system. This allows the estimation of peak velocities and wall-shear stress influencing the cultured cells in our multi-organ-chip (MOC) with respect to the applied pressure and the geometric properties of the pump. By adjusting those parameters, one can imitate physiological or pathological heart activity. The calculated flow within the MOC was compared to experimental results obtained via the non-invasive micro-PIV method.

scholarly journals The effect of insulin on equine lamellar basal epithelial cells mediated by the insulin-like growth factor-1 receptor

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e5945 ◽  
Author(s):  
Courtnay L. Baskerville ◽  
Subu Chockalingham ◽  
Patricia A. Harris ◽  
Simon R. Bailey
Keyword(s):  
Cell Proliferation ◽  
Growth Factor ◽  
Epithelial Cells ◽  
Cultured Cells ◽  
Maximal Effect ◽  
Insulin Like Growth Factor ◽  
Tissue Sections ◽  
Proliferative Effect ◽  
Increased Risk ◽  
A Cell

Background In horses and ponies, insulin dysregulation leading to hyperinsulinemia may be associated with increased risk of laminitis, and prolonged infusion of insulin can induce the condition. It is unclear whether insulin may have a direct or indirect effect on the lamellar tissues. Insulin is structurally related to insulin-like growth factor (IGF-1), and can bind the IGF-1 receptor, albeit at a lower affinity than IGF-1. Methods Immunohistochemistry was performed on formalin-fixed lamellar tissue sections from six normal horses, euthanised for non-research purposes, using an anti-IGF-1 receptor antibody. In further studies, lamellar epithelial cells were obtained by collagenase digestion from the hooves of 18 normal horses, also euthanised for non-research purposes, and incubated for 48 h in the presence of insulin (0–2,000 m IU/ml). The increase in cell numbers was determined using a cell proliferation assay, and compared to the effect of zero insulin using one-way ANOVA. Results Immunohistochemistry demonstrated IGF-1 receptors on lamellar epidermal epithelial cells. With cultured cells, insulin caused a concentration-dependent increase in cell proliferation compared to untreated cells (maximal effect 63.3 ± 12.8% more cells after 48 h with 1,000 m IU/ml insulin; P < 0.01). Co-incubation with a blocking antibody against the IGF-1 receptor significantly inhibited the proliferative effect of insulin (P < 0.01). Discussion These results demonstrate that IGF-1 receptors are present on lamellar epithelial cells. At high physiological concentrations, insulin may activate these cells, by a mechanism involving IGF-1 receptors, resulting in a proliferative effect. This mechanism could help to explain the link between hyperinsulinemia and laminitis.

scholarly journals Quantitative Analysis of Plasmodesmata Permeability using Cultured Tobacco BY-2 Cells Entrapped in Microfluidic Chips

2021 ◽  
Author(s):  
Kazunori Shimizu ◽  
Masahiro Kikkawa ◽  
Ryo Tabata ◽  
Daisuke Kurihara ◽  
Ken-ichi Kurotani ◽  
...  
Keyword(s):  
Real Time ◽  
Microfluidic Device ◽  
Cultured Cells ◽  
Cell Movement ◽  
Flow Channel ◽  
Microfluidic Chips ◽  
Isolated Cells ◽  
Cell Clusters ◽  
A Cell ◽  
Channel Structures

AbstractPlasmodesmata are unique channel structures in plants that link the fluid cytoplasm between adjacent cells. Plants have evolved these microchannels to allow trafficking of nutritious substances as well as signaling molecules for intercellular communication. However, tracking the behavior of plasmodesmata in real time is difficult because they are located inside tissues. Hence, we developed a microfluidic device that traps cultured cells and fixes their positions to allow testing of plasmodesmata permeability. The device has 112 tandemly aligned trap zones in the flow channel. Cells of the tobacco line BY-2 were cultured for 7 days and filtered using a sieve and a cell strainer before use to isolate short cell clusters consisting of only a few cells. The isolated cells were introduced into the flow channel, resulting in entrapment of cell clusters at 25 out of 112 trap zones (22.3%). Plasmodesmata permeability was tested from 1 to 4 days after trapping the cells. During this period, the cell numbers increased through cell division. Fluorescence recovery after photobleaching experiments using a transgenic marker line expressing nuclear-localized H2B-GFP demonstrated that cell-to-cell movement of H2B-GFP protein occurred within 200 min of photobleaching. The transport of H2B-GFP protein was not observed when sodium chloride, a compound known to cause plasmodesmata closure, was present in the microfluid channel. Thus, this microfluidic device and one-dimensional plant cell samples allowed us to observe plasmodesmata behavior in real time under controllable conditions.

Sign in / Sign up

Export Citation Format

Share Document