Trypan blue exclusion

Production of Mucosal Damage 2.3.1.2.1 Cell culture Stimulated neutrophils are known to be cytotoxic to cells in vitro (Dull et al., 1987 Dallegri et al., 1990 Grisham et al., 1990b). Several in vitro systems have been used to demonstrate oxidative damage to intestinal cells. Xanthine/XO increased Cr release and decreased [ H]thymidine uptake by IEC-18 small intestinal epithelial cell monolayers in a dose-dependent manner (Ma et al., 1991). Rat enterocytes show decreased trypan blue exclusion and increased protein release when incubated with neutrophils stimulated... 

Hwang and Bowen (2004) LNCaP Roche 10% water miscible beadlets 0.1-5.0pM DMSO in H20 6, 24, 48 h Lycopene uptake Cell viability by Trypan blue exclusion Cell cycle by flow cytometry... 

Trypan Blue exclusion and viable cell counts Cell viability by MTT... 

Trypan Blue exclusion (TB) Dead cells blue stained Cell membrane Yes3 [34]... 

MTX interferes with the growth of cancer cells by inhibiting the metabolism of folic acid. Drug efficacy was evaluated in vitro by MTT assay, as described above, and by Trypan Blue exclusion. Trypan Blue is a non-vital dye excluded by viable cells, but selectively staining dead cells. According to Figure 13.7, higher suppression of cell... 

Fig. 13.7 Effect of MTX-LDH hybrid on cell proliferation (A) and viability (B) as determined by MTT assay and Trypan Blue exclusion, respectively. HOS cells were incubated with free MTX, MTX-LDH hybrid or LDH for 72 h.

For purposes of quality control, any cell preparation used in the screen (see below) needs to have a definitive or consensus diagnosis and grading determined by histological examination. Furthermore, tumor cell preparations must be at least 80% tumor cells (as determined by cytopathological examination), whereas normal cell preparations must be devoid of any tumor cells. Cell viability, determined by trypan-blue exclusion, must be a minimum of 70%, and the signal-to-noise ratios for a predetermined cell cluster concentration must be at least threefold. 

Count viable cells by Trypan blue exclusion and seed cells at the appropriate density in culture medium in untreated tissue flasks. 

Count the cells by trypan blue exclusion test in a Biirker-Tiirk cell counting chamber and calculate the correct dilution (see below). 

Take a sample of the hollow-fiber product, determine cell viability by trypan blue exclusion (see Note 28), and assess the glucose level using the BM-Test 1-44 strips (see Note 26). 

Wash cells twice with labeling buffer by centrifugating at 300g and proceed to determine cell concentration using a particle counter. Also, perform a viability test using a Trypan blue exclusion method. Keep cells in cold storage (4°C) until used. 

Report cell population viabilities as measured by the Trypan blue exclusion test in the original and sorted cell samples. 

Prolonged and unnecessary enzymatic treatment, i.e., trypsinization of tumor cells, can also alter their survival and metastatic behavior in vivo. Moreover, viability tests (trypan blue exclusion) and even plating efficiency in vitro do not predict or correlate with the in vivo biological behavior of trypsinized cells. 

Viability of the hepatocytes is usually determined by trypan blue exclusion rate. Viability over incubation time can be determined by LDH retention or albumin secretion (Gebhardt 2003). For metabolism... 

C. Hepatocytes are counted in a hemocytometer and viability is determined using the trypan blue exclusion method. Viability is always > 90 % with a yield of 2-3 x 10s cells. Cells are resuspended in medium and diluted to a final concteration of 1 x 106 cells/ml. 

After an appropriate growth period (e.g. 4-6 days, depending on cell line, for serum batch testing), determine the endpoint (as below). For toxicity tests, incubate cells in 100 xl of medium for 24 h prior to addition of test substance (at twice the final concentration in 100 xl of medium). Incubation with toxic substance prior to endpoint measurement is usually for 6 days. For measurement of Trypan blue exclusion, trypsinize cells with 0.5% trypsin (Gibco) and... 

PRELIMINARY PROCEDURE TRYPAN BLUE EXCLUSION METHOD FOR CELL VIABILITY ESTIMATION... 

During the culture, the concentrations of living and dead cells (by the Trypan blue exclusion method) were measured using a haemocytometer (Chapter 2, section 2.2) glucose, lactate and glutamine by enzymic methods ammonia with a selective electrode and monoclonal antibodies by ELISA assay. 

Remove 1 ml of cell suspension into a disposable culture tube for a cell count. Determine the cell concentration of the suspension by direct counting and monitor the viability using the Trypan blue exclusion test. 

A sample is taken for counting (the trypan blue exclusion technique can be used to identify the percentage of dead cells) (see Note 6). 

Dead cells, as defined by Trypan blue exclusion, have been found not to contribute to the glutathione measurement. 

Maintain the cells in exponential growth by seeding at a density of 4 x 105-6 x 105 cells/mL, and count daily until a density of 2 x 106 cells/mL is reached. During this interval the cells are growing in a strict exponential fashion. In practice, the cells are subcultured and maintained with fresh medium containing 10% serum every 3 d. The cell viability should be maintained at approx 90-95%, as determined by the trypan blue exclusion method. 

Count the cell numbers and determine the viability by Trypan blue exclusion method. 

Trypan Blue Exclusion Test for Cell Viability... 

For statistical analysis, the greater the number of counts per culture the more accurate will be the statistics. The absolute minimum is triplicate values. Figure 2 shows the selective toxicity of EF13 for HIV-chronically infected cells over uninfected cells established using the Trypan blue exclusion test. 

The toxicity is evaluated by trypan blue exclusion method. The percent toxicity... 

We further tested this effect by inducing " " chemical hypoxia (20, 21). Potassium cyanide (KCN, 2.5 mM) was used in endothelial culture for 2 h. In the study group (JST=6), ATP-vesicles were added while no ATP-vesicles were added into the control group (N=6). After 2 h, the cells in the study group maintained more than 90% viability as determined by trypan blue exclusion. In the control, no more than 10% of the cells were still alive (Fig. 3). 

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