Monolayer cell culture modifications

The test of inflammatory reactions consisted of quantifying the MGC in monolayer cell cultures of L132 cells. This test reveals morphological modifications in a cell culture by the appearance of MGC, which are directly... 

In the selection of an appropriate cell culture system, a number of criteria must be considered (Table 3). These include not only the characteristics of the cell type but also the controllable parameters of the complete transport system such as the permeants, the filter properties, and the assay conditions. In general, most transport experiments employ the experimental design shown schematically in Figure 4 with modifications as discussed below. Typically, the desired cell is seeded onto some sort of semipermeable filter support and allowed to reach confluence. The filter containing the cell monolayer separates the donor and receiver... 

The cultivation of hepatocytes in a stationary suspension culture is actually ineffective. The hepatocytes lose their differentiation within hours. An improvement was the attachment culture. Thereby, the cells are cultivated either in self- or microcarrier-induced multicellular aggregates or on membranes. When standard monolayer culture was adequate to maintain the cell viability for 1 to 2 weeks the differentiation was lost after a few days. Different modifications as described beneath allowed the maintenance of differentiation for 2 to 3 weeks. 

Aside from encapsulation of proteins, much work has been done on the surface functionalization of microparticles for cellular interaction and proliferative effects. Surface modification of PLGA microspheres with an amine-terminated dendrimer improved long-term proliferation of chondrocytes without observed changes in the cell phenotype, as compared to monolayer culture systems (Thissen et al. 2006). Additionally, surface functionalization ofpolystyrene magnetic microbeads with the DLL4 notch ligand used in coculture has been shown to efficiently generate T cells from mouse bone marrow hematopoietic stem cells (Taqvi et al. 2006). 

For studies of anchored cells, modifications need to be made in many cases. For instance, cultures on tenterframes [35] have been inserted into the lOO-cm batch vessel of a Calvet calorimeter [3] and glass plates with cell monolayers on them have been stacked in the ampoules of a Thermometric BAM instrument [36]. In one recent case [37], the stirrer blades of a Thermometric perfusion vessel were modified to act as monolayer plates. It would seem preferable these days to attach the cells to beads that are suspended in a stirred ampoule (see for instance Reference [38]). 

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