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FIGURE 7.1 Morphology on Feeder Layers. Morphological alteration in cells growing on feeder layers: Fibroblasts from human breast carcinoma (a) growing on plastic and (b) growing on a confluent feeder layer of fetal human intestinal cells (FHI). Epithelial cells from human breast carcinoma growing (c) on plastic and (d) on same confluent feeder layer as in (b).
FIGURE 7.2 Cell Yield and Surface Area. Relationship of volume of medium and cell yield to the surface area of a culture vessel. The graph is plotted on the basis of the volume of the medium for each size of vessel and is nonlinear, as smaller vessels tend to be used with proportionally more medium than is used with larger vessels. The cell yield is based on the volume of the medium and is approximate.
FIGURE 7.3 Multiwell Plates. Six-well, 24-well, and 96-well (microtitration) plates. Plates are available with a wide range in the number of wells, from 4 to 144 (see Table 7.2 for sizes and capacities).
FIGURE 7.4 Petri Dishes. Dishes of 3.5-, 5-, and 9-cm diameter. Square Petri dishes are also available, with dimensions 9 × 9 cm. A grid pattern can be provided to help in scanning the dish—for example, in counting colonies—but can interfere with automatic colony counting.
FIGURE 7.5 Plastic Flasks. Sizes illustrated are 10 and 25 cm2 (Falcon, BD Biosciences), 75 cm2 (Corning), and 185 cm2 (Nalge Nunc) (see Table 7.2 for representative sizes and capacities).
FIGURE 7.6 Multisurface Flask. The Nunc Triple-Flask with three 80-cm2 growth surfaces that are seeded simultaneously. Although the growth surface is 240 cm2, the shelf space is equivalent to a regular 80-cm2 flask (See also Corning HyperFlask, Fig. 26.9). As the head space for gas phase is smaller, this flask is best used with a filter cap in a CO2 incubator (arrow). (Photograph courtesy of Nalge Nunc International.)
FIGURE 7.7 Stirrer Flasks. Four small stirrer flasks (Techne), 500-mL capacity, with 250 mL medium, on four-place stirrer rack (see also Figs. 12.5, 26.1, 26.2). (Courtesy of Sterilin.)
° FIGURE 7.8 Venting Petri Dishes and Flasks. (a) Vented Petri dish. Small ridges, 120 apart, raise the lid from the base and prevent a thin film of liquid (e.g., condensate) from sealing the lid and reducing the rate of gas exchange. (b) Gas-permeable cap on 10-cm2 flask (Falcon, BD Biosciences).
FIGURE 7.9 Screw-Cap Vials and Flasks. (a) Glass flasks are suitable for replicate cultures or storage of samples, particularly when plastic may not survive downstream processing. Screw caps are preferable to stoppers, as they are less likely to leak, and they protect the neck of the flask from contamination. (b) Scintillation vials are particularly useful for isotope incorporation studies but should not be reused for culture after containing scintillation fluid.
FIGURE 7.10 Nonrandom Growth. Examples of ridges seen in cultured monolayers in dishes and flasks, probably due to resonance in the incubator from fan motors or to opening and closing of the incubator doors. (Courtesy of Nunc.)
FIGURE 7.11 Hollow Fiber Culture. A bundle of hollow fibers of permeable plastic is enclosed in a transparent plastic outer chamber and is accessible via either of the two side arms for seeding cells and collecting high molecular weight product. During culture the chamber is perfused from a reservoir, via a gas exchanger and peristaltic pump, down the center of the hollow fibers through connections attached to either end of the chamber. (Courtesy of FiberCell, Inc.; see also Fig. 26.6; Plate 24.)
