Cell detachment

No spermatozoa are seen on the egg surface or on the vitelline coat. After 5 h of treatment with 10 M TBTCl, a few spermatozoa, with very anomalous heads, have been detected. The absence of spermatozoa on the egg surface or on the vitelline coat could be explained by the absence of the follicle cells, which, in S. plicata, primarily play an attracting function. It was previously shown that TBTCl solution, either 10 or 10 M, induces anomalies in spermatozoa, unfertilized, and fertilized eggs of Ascidia malaca. In particular, the follicle cells detach from eggs and the test cells show anomalies in their nucleus and granules. Moreover, damaged spermatozoa are observed in the vitelline coat, but never in... 

In some studies it has been shown that ITCs can cause increases in the pro-apoptotic caspase enzymes, caspase 3, caspase 8 or caspase However, in other work, specific caspase inhibitors failed to block cell death or cell detachment from the substratum . This suggests that caspase-activation may only be a bystander event, or may only occur after the initiating event of a block in the cell cycle. Indeed, although expression of c-Jun amino-terminal kinase (INK) in response to ITCs has been linked to the pro-apoptotic process, it remains entirely possible that this is a parallel signalling pathway, more closely related to the induction of Phase 1 or Phase 2 enzymes than to the... 

From Table 4 it is evident that several natural and modified polyanions did not induce insulinoma cell detachment. Furthermore, the DNA content, a quantitative metric of cell viability, was in general near the baseline (100%) observed in the control studies with PBS. In generalboth the cell attachment and DNA indices gave similar indications of cytotoxicity. Therefore, in an effort to unify these measures the third column of Table 4 reports an overall ranking of the polymer... 

Okano, T., Yamada, N., Okuhara, M., Sakai, H., and Sakurai, Y. Mechanism of cell detachment from temperature-modulated, hydrophilic-hydrophobic polymer surfaces, Biomaterials, 1995, 16, 297-303. 

Harvest the cells by ttypsinization. Trypsinization is accomplished by decanting the medium from a flask of cells and adding 5 mL of trypsin-EDTA. Wait until the cells detach (approx 5 min), then ttansfer the cell suspension to a centrifuge tube. Pellet the cells by centtifugation at 500g for 5 min. 

Cells detachment with trypsin is performed at room temperature and not at 37°C as usually indicated. This is needed to reduce the number of movements and hence to decrease the risk for the cylinder to detach from the surface with the consequent loss of the colony. 

Okano, T., N. Yamada, M. Okuhara, H. Sakai, and Y. Sakurai. 1995. Mechanism of cell detachment from temperature-modulated hydrophilie-hydrophobie polymer surffitematerialsl6 297-303. 

Grow the HT-29 cell line in T75 flasks in McCoy s 5A medium (10% FBS). When cells are confluent, harvest HT-29 cells by adding 2.0 ml of TrypLE Express. After the cells detach from the flasks, resuspend the cells in McCoy s medium (10% FBS) to inactivate the TrypLE Express. Count the cell concentration as described below. Prepare two small T25 flasks with McCoy s medium (10% FBS) and seed 1 x 106 cells per flask. Keep the HT-29 cells in culture as they will be needed again later. 

Check for cell detachment by holding unit up to the light. Return unit to hood, remove cap, and pour off cell suspension into the collection flask containing FBS. 

On the other hand, multiple laminar flow was used to pattern cells and their environments in PDMS chips [858]. For instance, two cell types (chicken erythrocyte and Escherichia coli) have been shown to deposit next to each other on fibronectin-treated surfaces (see Figure 8.17). Moreover, cell detachment occurred only to the cells (BCE) that was passed with a patterned stream containing trypsin/EDTA [858]. 

Bursts of HA deposition correlate with mitosis.41 13 Elevated levels promote cell detachment, in preparation for mitosis, as cells leave tissue organization, and enter the transient autonomy required for the mitotic event to occur. Cells must then degrade that HA after mitosis has occurred, to regain adhesiveness, and to reenter the social contract. The prediction is that HA synthesis occurs as... 

Levels of HA that cells deposit must respond to various physiological states including growth phase,239 confluence, inversely related to cell density in both fibroblasts,240 and keratinocytes,241 mitosis and cell detachment from the substratum,242 calcium concentrations,149,243 anoxia and lactate,244 viral transformation,245 and serum stimulation.91,246 Preliminary immunolocalization data indicate that some of the HAS and hyaluronidases colocalize (A. Spicer et al., pers. commun.). All of this evidence supports, albeit indirect and tentative, the existence of the hyaluronasome structure. 

S.C.J. Reader, and R.M.D. Foster, The in vitro effects of four isomers of dinitro-toluene on rat sertoli and sertoli-germ cell co-cultures Germ cell detachment and lactate and pyruvate production. Toxicol. Appl. Pharmacol. 106 287-294, 1990. 

Recently, TTR1 fibrils have been decorated with the classic RGD tripeptide motif isolated from fibronectin, which encourages cell adhesion via integrin cell surface receptors (Gras et al., 2008). This bioactive tag was added to the TTR1 sequence which drives fibril assembly and the tag shown to be exposed on the fibril surface and accessible to cells following assembly. The RGD-modified fibrils were bioactive in a cell dissociation assay which measures the ability of fibrils to competitively bind to cells and induce cell detachment from a surface, as illustrated in Figure 19. In... 

Add 150 pL trypsin solution to each well until the cells detach. 

Protein content Cell growth, cell detachment Easy to measure... 

Robinson BW, Venaille TJ, Mendis AH, McAleer R Allergens as proteases An A. fumigatus proteinase directly induces human epithelial cell detachment. J Allergy Clin Immunol 1990 86 ... 

Tomee JF, Wierenga AT J, Hiemstra PS, KauSman HE Proteases from A. fumigatus induce release of proinflammatory cytokines and cell detachment in airway epithelial cell lines. J Infect Dis 1997 176 300-303. 

Giant and atypical mesothelial cells can develop in the peritoneum in response to irritation caused by peritoneal dialysis solutions. Mesothelial hyperplasia without an increase in mesothelial cell size, cytological atypia in mesothelial cells, polynucleate cells with nucleoles, and enlarged mesothelial cells with a flat small nucleus and without evident nucleoles have all been described (5). These giant cells are not found unless there is peritonitis. During peritonitis mesothelial cells detach en masse from the basal lamina. 

Racusen EC, Fivush BA, LI YL, Slatnic I, Solez K Dissociation of tubular cell detachment and tubular cell death in clinical and experimental "acute renal failure". Lab.Invest. 64 546-556,1991... 

Cells are cultured in Eagle s MEM supplemented with 10% horse serum or FBS. The cells detach at room temperature and may take days to reattach. 

Harvest cells when confluent (5-6 days) by removing the medium and trypsinizing in the conventional way. After adding trypsin, roll the bottles at speeds of 20-60 rph until cells detach. Cell yields will be similar to or up to two-fold higher than in stationary cultures, and multilayering (non-diploid cell lines) will occur. 

Figure 10.4 (a) Resistance of bovine tracheai mucosa to injury eiicited by ionophore activated human neutrophils, and (b) susceptibility of bronchial mucosa under identical conditions. Injury, In this case cell detachment, was measured indirectly by measuring the permeability of the airway mucosa to serum albumin. Cells were applied to the basolateral surface and the vector of solute movement was basolateral-apical. 

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