Cell Culture High-Rise

A little over a year ago, BioCrystal of Westerville, Ohio, introduced the futuristically designed OptiCell™ for tissue culture.1 The 100-cm2 growth area of each OptiCell (33% more than a T-75 flask) requires only 10 ml of growth medium. Building on this invention, BioCrystal developed a fully automated Robotic Cell Culture System (RC2S) capable of processing 402 OptiCells from inoculation to harvest with almost no human intervention. "Our focus is on growing anchorage-dependent cells," says Jeff Bergen, CEO. "Most pharmaceutical companies want to be able to harvest up to one billion cells per day on consecutive days for a week at a time for further processing;" continues Bergen. "Our RC2S system will enable that at reasonable cost." The current estimated price is about $400,000 (US). Tools for bulk cell culture currently include stir-tank ($150,000-$200,000) and cell-cube ($50,000-$100,000) bioreactors, roller bottles, and hollow-fiber systems. "A roller bottle output of 8-10 billion cells (cell-line dependent) would require at least 15 liters of medium, assuming no medium change was necessary," comments Ed McKillip senior research scientist and supervisor of the bioreactor labs in Princeton, NJ, for Radnor, Pa.-based Wyeth. An equivalent output can be achieved with only 10 ml of medium per cassette in the RC2S, thereby using four liters of the most costly reagent needed, or about 25% of a roller bottle run. After reviewing the specifications of the RC2S, McKillip adds that having multiple reagent delivery lines (currently 10) means that researchers can expose a single cell line to many different test compounds, or treat many different cell lines with one reagent. Unlike conventional cell culture that entails nonsterile trips in and out of the incubator, biosafety cabinet, and centrifuge, the RC2S is self-contained, HEPA filtered, and UV sterilized, and it provides continuous bioreagent filtration. The "do not disturb" sign on the incubator door would be ignored only during routine maintenance. "Temperature and humidity remain constant by doing everything in an incubation-type domain all the time and never moving materials out of that [environment]," says Bergen. The contiguous reagent cabinet offers temperature-controlled reagent management under filtered laminar airflow, and the clean incubator area houses a centrifuge for sample concentration. Researchers must optimize each cell line for growth in the OptiCell, says Chris Wolford, applications specialist at BioCrystal. They then program the bar-coded OptiCells' stay in what Bergen calls the "carousel motel," and the robot takes over. The robot reads the barcode and then processes the inserted OptiCells. The operator can program OptiCell flipping (to seed both sides), reagent changes, sample insertion or withdrawal, three shaking speeds, trypsinization, centrifugation, and culture expansion into additional units. "If researchers are looking for mammalian cells to support their high-throughput screening needs," comments Scott Waniger, cell culture production manager at the Minneapolis-based National Cell Culture Center (NCCC), "the RC2S would fit the bill." Because NCCC customers typically require 100 to 1,000 times as many cells as the OptiCell unit's maximum output, however, Waniger says the RC2S wouldn't be useful for his large-scale needs. BioCrystal is presently demonstrating the RC2S to prospective buyers to obtain feedback for final tweaking of the system. "The hard work's been done," says Bergen. The company expects to receive orders by midyear for delivery by year's end, he says. --Susan Jenkins (Note: the author worked at Wyeth from 1994 to 1999) 1. J.M. Pisano, "Tissue cultural revolution," The Scientist, 15[7]:22, April 2, 2001. For More Information BioCrystal +1 (614) 818-0019 www.opticell.com

Cell Culture High-Rise

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