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Inoculation, Growth & Harvesting in a New Easy-to-Use Disposable Cell Culture System
Sponsored,vendor-submitted protocol    Published in November 2006 (p.15) DOI: 10.2144/000112364

Introduction

New Brunswick Scientific now offers a new labor-saving device for producing mg quantities of proteins, virus or cell mass from anchorage-dependent or suspension cultures. More productive than traditional roller bottles, spinners or T-flasks, the new FibraStage system is inexpensive, simple to operate, and disposable. Typical yields of 4.9 × 109 cells per 500 mL disposable FibraStage bottle have been achieved with a variety of cells types. The system holds up to four bottles, enabling simultaneous screening of four different cell lines or media compositions.

At the heart of the system are pre-sterilized, single-use bottles, with a cell growth chamber filled with 10 grams of FibraCel disks. FibraCel is a three-dimensional growth substrate, composed of polypropylene and polyester non-woven fiber, which provides an extremely high-surface area for cell growth. Anchorage-dependent and suspension cultures become attached to or embedded in the FibraCel disk bed, where they remain throughout the process. Immobilization within the disk bed not only protects the cells from damage by shear forces, but also simplifies harvesting, allowing the supernatant to be easily recovered from the cell-free medium. The bottles come pre-sterilized and ready to use, requiring no cleaning, autoclaving or validating, for further reduction of labor.


Figure 1. (Click to enlarge)


The FibraStage system includes a variable speed device with movable platform into which the FibraStage bottles are placed. When the platform is raised, bellows at the base of each bottle are compressed, forcing media over the cells, facilitating nutrient uptake and removal of cell waste. When the platform is lowered, the bellows expand, allowing the media to recede, leaving the disks covered by a thin liquid film. The film not only limits damage to the cells, but helps with oxygen transfer and removal of excess CO2. The entire device fits into a standard CO2 incubator for control of temperature and gasses, and the controller is magnetized for convenient storage on the outside wall of the incubator.

FibraCel disks have a proven record of high-yield production of cells, virus and secreted proteins from a wide variety of cell lines, including hybridoma, CHO, VERO, BHK, and insect cells(1). These disks are also currently being used in the production of the world's first licensed gene therapy drug, a treatment for head and neck squamous cell carcinoma. New Brunswick Scientific offers a wide range of bioreactor systems with packed-bed basket technology in 0.5 to 150 Liter capacities (working volume), but FibraStage is the first of their single-use systems for cell culture.

Inoculation

Begin with an inoculum of 1-2 × 108 cells per FibraStage bottle for mammalian cell cultures, and 1.5-2×108 cells per bottle for insect cell cultures. Higher cell densities will shorten the lag phase of the culture.

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