Plastic flask

Cells in vivo exist either attached to a surface or free in suspension. Adherent cell lines originate from cells of solid tissue. Breast carcinoma cell lines (such as MCF7, T47D, and SK-BR-3) are adherent cultures, and these cells are grown on the surface of plastic flasks that have been treated to facilitate adhesion (see Fig. 6.2). Suspension culture cell lines originate from cells that exist in suspension, such as those cells present in the blood and the lymphatic system (see Fig. 6.3). 

The cooled colloid is filled into and stored in a plastic flask cleaned with Soln. D. 

Centrifuge these cell solutions for 5 min at 700 rpm and resuspend the pellet in culture medium for the 1st day. Seed the cells in 25-cm2 plastic flasks and incubate at 37 °C/5% C02. After 2 h of preplating (this time is required for nonmuscular cells to attach) the supernatant of this flask is filtered through a nylon mesh (pore width 100 pm) and then seeded in Petri dishes at a density of about 10,000 to 100,000 cells/cm2 in culture medium for the 1st day. After 24 h the Petri dishes are rinsed off with Dulbecco s wash solution and culture medium for the 1st... 

Cell Culture. Human neuroblastoma IMR-32 (passaged through nude mice the cells were donated by Dr. Steven E. Brooks, Kingsbrook Jewish Medical Center, Brooklyn) and mouse neuroblastoma clones NIE-115, NS-20, and N-18) (donated by Dr. Shraga Makover, Hoffmann LaRoche, Inc., Nutley, New Jersey) were maintained in our laboratory as described previously (30,31). Confluent monolayers (6 to 8 x 10 cells per 250-ml Falcon plastic flask) were harvested for enzymatic studies with phosphate-buffered saline [7.0 mM potassium phosphate/0.14 M NaCl - buffer, pH 7.2 (Pi/NaCl)] containing 0.1% EDTA. 

Depending upon the design of an experiment, cells were grown in 60 mm petri dishes, 150 cm2 plastic flasks (Falcon no. 3024), or roller bottles in MEM supplemented with 12% fetal bovine serum and antibiotics in 95% air, 5% C02 water humidified incubator at 37°. All experiments were initiated with confluent mono-layer of cells grown for about 5-15 generations. 

Cells may be grown in dishes or flasks where the initial inoculum varies from 0.2 X 106 up to 2 X 106. The containers may be glass or plastic. The plastic ware is obtained in sterile wraps from commercial suppliers and is specially prepared for use in cell culture (Fig. 3.2a) (see Appendix 3). The glass bottles are usually medical flat bottles but any bottle with a flat side will do provided it is washed correctly and sterilised before use (see Chapter 8). To a large extent the disposable plastic flask has now replaced the glass bottle. 

NovikofTcells were seeded in a plastic flask with a cell count of 0.1 x 10 cells. The compounds were dissolved in DMSO, diluted with the same volume of Hanks buffer and added to the culture medium. Assigned concentration is the end concentration of the compound in the liquid culture medium. Cell cultures were incubated for 4 days, and the vital cells were counted under a light microscope. The IC50 value represents the average of N = 6. Untreated cell culture served as control. 

A solution of DAST (1 0.484 g, 3.0 mmol) in CHjClj (2 mL) was placed in a plastic flask and cooled to — 80 C. A solution of ethyl 2-hydroxy-2-phcnylpropanoate (0.291 g, 1.5 mmol) in CHjClj (2mL) was added drop wise with stirring over a period of 10 min under argon. The whole mixture was allowed to warm to rt. stirred at rt for 2 h, poured into H O (30 mL), and extracted with CHjClj (3 x 10 mL). The combined extracts were washed with sat. brine and dried (MgSO ). Evaporation of the solvent gave a crude product, which was purified by chromatography (silica gel) to give a pale-yellow oil in quantitative yield. 

HSV (type 1 or 2) stock is prepared by infecting Vero cells (Id after passage at 1 4) in plastic flasks at 37°C. Check by microscopy for cytopathic effect... 

For long-term storage, it is recommended that plastic flasks be used. 

Tissue culture-treated plastic flasks, dishes, and 24-well plates. 

Dissolve 400 g of reagent purity sodium hydroxide and purity potassium sodium, tartrate in dist. water, cooling make up to 1 litre. Store the solution in a plastic flask. 

Next rinse out the plastic flask with about 20 ml of concentrated reagent-... 

All solutions used must be stored in plastic flasks so as to ensure that no silicic acid is dissolved out from glass. 

Spectrophotometer with 1-cm and 5-cm cuvettes Plastic flasks and beakers, 100 ml... 

In order to remove any last traces of Si02 from the distilled water to be used for silicic acid determination, run the water through a column with a strongly basic anion exchanger (OH"-type). The column should be used for the discharge piping. Store the silicic-acid-free water in plastic flasks. 

Introduce approximately 2 g 0.2 g of hydrofluoric acid/perchloric acid mixture into 100 ml plastic flasks. Pipette in 50 ml of the water sample, mix and leave to stand for a few hours. If the plastic flasks are charged with the decomposition acid and weighed beforehand, the water sample may also be filled into the flask directly at the sampling point. The transport time to the laboratory is then used for the acid decomposition of the... 

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