Corning® Roller Bottles Selection and Use Guide Innovative Products for Biotechnology Table of Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 Corning Disposable Plastic Roller Bottles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 Corning PYREX® Glass Reusable Roller Bottles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 Tips on Solving Growth Problems in Roller Bottles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 Technical Assistance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

Introduction In the past 90 years, animal cell culture has evolved from a laboratory curiosity into one of the leading tools of biotechnology. During this time the products used for growing cells, especially culture vessels, have also undergone an evolution. This is especially true of roller bottles. The concept of growing cells as rotating cultures was originally developed by George Gey (1933) at Johns Hopkins University as a means of growing larger quantities of anchorage(attachment) dependent cells. His work was primarily done in glass roller tubes. By the 1960s, much larger glass roller bottles...

Standard Corning® polystyrene roller bottles are surface modified for improved cell attach- ment using a traditional gas-plasma treatment. This generates highly energetic oxygen ions which graft onto the surface polystyrene chains so that the bottle surface becomes hydro- philic and negatively charged so cells can better attach.. Polystyrene roller bottles are also available with a Corning CellBIND® Surface treatment that uses a novel microwave plasma process for treating the culture surface. This process improves cell attachment by incorpo- rating significantly more oxygen into the cell culture...

Corning® Expanded Surface Polystyrene Roller Bottles Although these bottles have the same approximate outer dimensions of the 850 cm2 roller bottles, the ribbed design provides twice the growth surface while maintaining the same exterior dimensions. Two smooth window areas that run the length of the bottle provide microscopic viewing as well as pouring surfaces. The bottles are graduated from 100 to ► The standard Corning tissue culture treated surface ► The Corning CellBIND surface treatment ► Easy Grip caps are designed for more comfortable manual handling, have half turn on/off seating, smooth,...

PYREX Glass Roller Bottles with 45 mm Screw Cap Bottles have linerless, one-piece orange polypropylene plug seal caps with GL45 threads and drip-free pouring rings. A wide range of optional caps are available including: colored caps for ease of sorting and identification or vented membrane caps with 0.22 µm PTFE hydrophobic membranes for gas exchange. Corning Cat. No. Cell Growth Area (cm2) Roller Bottle Roller Bottle Roller Bottle Roller Bottle PYREX Glass Roller Bottle with 51 mm Screw Cap Wider mouth simplifies cell harvesting. Bottles are supplied with 51 mm phenolic, rubberlined caps. Corning...

Another possible solution to cell attachment problems in roller bottles is the Corning® CellBIND® Surface. The first novel cell culture surface treatment in over 20 years, this surface is designed to improve cell attachment under difficult conditions, such as growth roller bottles. It is also useful for growing “difficult” cells such as primary cultures or transfected cells over expressing proteins. Developed by Corning scientists, this technology uses a microwave plasma process for treating the culture surface. This process improves cell attachment by incorporating significantly more oxygen...

Clear Bands Figure 3. Two clear bands caused by debris in the medium scraping away cells as the bottle was rotated. Occasionally clear circular bands will occur on roller bottles where the cells appear to have been swept away (See Figure 3). While small pieces of rolling debris or large cell clumps can cause this to occur, one of the most common causes is the short-term presence of bubbles in the initial cell inoculum. These bubbles, when in contact with the sides of the slowly rotating bottle, can act as miniature plows, scraping off the cells as they begin to attach. Avoid bubble formation...

Poor Growth Many problems with the growth, maintenance, or performance of cell cultures are eventually traced back to the medium in which the cells are cultured. These medium-related problems are often slow to develop and are rarely obvious, usually requiring significant effort to track down and eliminate. Unless heavily contaminated, good culture medium is not visibly differ- ent in appearance from defective culture medium. The only good way to determine medium quality is to attempt to grow cells with it; this is the basic quality control procedure used by reliable commercial media manufacturers....

Fluorescent Light-induced Toxicity The deleterious effect of fluorescent light on culture media may be the single most overlooked source of chemically induced cytotoxicity. It is very important to store media and cells growing in culture vessels in the dark away from sources of fluorescent light that will interact with light sensitive media components (riboflavin, tryptophan and HEPES). These interactions result in the production of hydrogen peroxide and free radicals that are directly toxic to cells. This well-documented problem is often ignored when there are cell growth issues (Wang, 1976,...

Technical Assistance For additional technical support and product information, please call 1.800.492.1110 or 1.978.442.2200. Technical and product information is also available on the Corning Life Sciences web site at www.corning.com/lifesciences. References Freshney, R.I. (1994). Culture of animal cells: A manual of basic technique — 3rd edition. Wiley-Liss, Inc. New York, pp. 373-375. Gey, G.O. (1933). An improved technic for massive tissue culture. Amer. J. Cancer 17:752-756. Lincoln, C.K. and Gabridge, M.G. (1998). Cell culture contamination: sources, consequences, prevention and elimination....