Cell adherence

Cutting sections at 3 pm facilitates single cell adherence to charged slides. The presence of nonadhered cells sloughing off between cell spots is a key characteristic of thick cell line sections (see Fig. 6.8). An awareness of actual... 

It has been shown that cell adhesion highly depends on the outermost functional groups on SAMs however, cells do not directly interact with the SAMs. Instead, they interact with proteins adsorbed on SAMs. Cell adherence requires an interaction between integral molecules in the cell membrane and glycoproteins specialized for cell adhesion, like fibronectin (Fn) and vitronectin (Vn), which are adsorbed on the artificial material. Thus, the presence of glycoproteins in serum plays a crucial role in cell adherence to artificial materials. In the first part of this review (Sect. 2), we will briefly survey recent studies of cell adhesion on SAMs with different functional groups and discuss the mechanisms involved. 

When a cell suspension is applied to a surface, the events that occur can be conceptually classified into three stages (1) a cell approaches the surface, (2) the cell attaches to the surface, and (3) the cell adheres, and thus, spreads out on the surface. Most studies of cell adhesion on artificial materials measure the number of adherent cells, the cell morphology, and changes in protein expression. To gain more detailed insight into the biophysical mechanism of cell adhesion requires... 

TIRFM was used for time-lapse observations of initial cell adhesion to SAMs with different surface functionalities (Fig. 2). After 10 min of plating a suspension of human umbilical vein endothelial cells (HUVECs), a few bright spots were observed on SAMs with COOH and NH2 functionalities this indicated cell adherence. The number of bright spots increased and the spot areas enlarged with incubation time, indicating that HUVECs adhered and spread well on COOH-SAM and NH2-SAM surfaces. Quantitative analysis of the number of adherent cells and cell adhesion areas... 

Figure 4c shows one example of the time course of an SPR angle shift during exposure of a NH2-SAM to culture medium supplemented with 2% fetal bovine serum (FBS). It also includes the time course of the fraction of adherent cells on the same surface determined by TIRFM observation (Fig. 2). The SPR angle shift rapidly increased, and then leveled off within a few minutes. Cells adhered much more slowly than proteins. Those results indicated that serum proteins in a medium rapidly adsorbed to the surface then, cells interacted with the adsorbed protein layer, as shown schematically in Fig. 5.

Figure 4c shows that the amount of adsorbed proteins is rapidly saturated within several minutes of exposing serum-containing medium to a surface. Albumin, the most abundant serum protein, was expected to preferentially adsorb onto the surfaces during early time points. Then, adsorbed albumin was expected to be displaced by cell adhesion proteins. To investigate the effect of preadsorbed albumin displacement on cell adhesion, SAMs were first exposed to albumin then, HUVECs suspended in a serum-supplemented medium were added [21, 42]. Very few cells adhered to hydrophobic SAMs that had been pretreated with albumin, due to the large interfacial tension between water and the hydrophobic surfactant-like surface. Albumin was infrequently displaced by the cell adhesive proteins Fn and Vn. One the other hand, HUVECs adhered well to hydrophilic SAM surfaces that had been preadsorbed with albumin. In that case, the preadsorbed albumin was readily displaced by cell adhesive proteins. 

We investigated the efficiency of NSC expansion on surfaces with EGF-His immobilized in the correct orientation. NSCs were obtained from neurosphere cultures prepared from fetal rat striatum harvested on embryonic day 16. NSCs were cultured for 5 days on EGF-His-immobilized substrates prepared with mixed SAMs of different COOH-thiol contents. Cells adhered and formed network structures at a density that increased with the COOH-thiol content of the surface. As a control, cells were seeded onto surfaces without immobilized EGF-His. This resulted in poor cell adhesion during the entire culture period. In addition, when EGF-His adsorbed to SAMs with 100% COOH-thiol or SAMs with NTA-derivatized COOH that lacked Ni2+ chelation, we observed poor initial cell adhesion, and the cells formed aggregates within 5 days. Interestingly, the substrate used to covalently immobilize EGF-His with the standard carbodiimide chemistry was not a suitable surface for cell adhesion and proliferation. The control experimental results contrasted markedly with results from EGF-His-chelated surfaces. 

The application of fullerene on the surfaces has an essential advantage in the studies with cell cultures as in this case we can obtain the maximum contact of cells with fullerene - cells adhere on the surface and colonize it as a confluent monolayer. That is the basic difference from the water-soluble complexes and micro-dispersed suspensions of fullerene C60. The pro-/antioxidant activities of fullerene were tested in chemical and biological systems. 

Demling N, Ehrhardt C, Kasper M, Laue M, Knels L, Rieber EP (2006) Promotion of cell adherence and spreading a novel function of RAGE, the highly selective differentiation marker of human alveolar epithelial type I cells. Cell Tissue Res 323(3) 475-488... 

Tatsuno, I., Horie, M., Abe, H., Miki, T., Makino, K., Shinagawa, H., Taguchi, H., Kamiya, S., Hayashi, T., and Sasakawa, C. (2001). toxB gene on p0157 of enterohemorrhagic Escherichia coli 0157 47 is required for full epithelial cell adherence phenotype. Infect. Immun. 69, 6660-6669. 

If cellular localization of the antigen-antibody complex is not required, enzyme immunolabeling can be performed on cells adherent to a microtiter plate, and the color change resulting from the enzymatic reaction can be detected as a change in absorbance with an automatic plate reader (see Chapter 28). Biotinylation of antibodies and the use of the avidin-biotin complex has further extended the versatility and sensitivity of the enzymatic techniques (see Chapters 7 and 25-27). Most recently, the principles behind these techniques have been applied in combination with in situ hybridization techniques. Using nucleic acid-antibody complexes as probes, specific DNA or RNA sequences can be localized (see Chapters 46 9). 

Gambierdiscus toxlcus cells adhered to the Inside surface of... 

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