Development of a Low Capital Investment Reactor System: Application for Plant Cell Suspension Culture

Abstract

Growth of plant cell cultures is demonstrated in an uncontrolled, simple, and inexpensive plastic-lined vessel. Sustained specific growth rates of 0.22 day-1 for Hyoscyamus muticus cell suspension cultures are achieved in a low-cost gas-sparged bioreactor configuration (6.5 L working volume, wv) which is comparable to an "optimized" 5 L wv mechanically agitated fermentor. In an effort to reduce bioreactor costs, the need for an autoclavable vessel was eliminated. Sterilization is achieved by separate autoclaving of the plastic liner and by gas-phase sterilization using ethylene oxide. The initial run sterilized with ethylene oxide displayed a long lag, apparently due to residual sterilant gas. Because ethylene oxide could eliminate costs associated with autoclave rated vessels, a quantitative basis for aeration time was developed by experimental measurements and modeling of diffusion in the polymer liner. Operational techniques to eliminate toxicity are implemented to grow 0.2 kg dry weight of plant cells in 13 days in a 40 L (28.5 L wv) air-lift bioreactor without autoclave sterilization. The biomass yields for all reactors were statistically indistinguishable from shake flask culture.

Improvement of plastic-based disposable bioreactors for plant science needs

“…In order to decrease the production costs, a few alternatives to traditional stainless-steel bioreactors have recently been developed (Singh 1999;Hsiao et al 1999;Curtis 1999), but these systems, when used for plant cell cultures, concern only small working volumes (Palazon et al 2003;Bentebibel et al 2005;Eibl and Eibl 2006). We are developing two new flexible, scalable, plastic-made disposable bioreactors (Terrier et al 2007).…”

Two new disposable bioreactors for plant cell culture: The wave and undertow bioreactor and the slug bubble bioreactor

“…The running costs are high, due to low yields and the need to clean and sterilize the bioreactor after each culture cycle. In order to decrease the production costs, a few alternatives to traditional stainless-steel bioreactors have been developed (Curtis, 1999;Hsiao et al, 1999;Singh, 1999). Singh (1999) developed a disposable bioreactor with an original agitation apparatus, using an inflated bag placed on a rocking mechanism that moves the bag inducing a wave-like motion to the liquid contained therein.…”

Production of mouse interleukin‐12 is greater in tobacco hairy roots grown in a mist reactor than in an airlift reactor

“…Use of a plastic bag as a root growth chamber builds on concepts conceived by others (Hsiao et al, 1999;MedinaBolivar and Cramer, 2004) and enables creation of an artificially dense root bed inoculum which increases the probability of droplet capture and enhanced root growth. The Velcro constriction further provides not only a seat for the inoculum, but also aids in funneling the bag wall thereby clustering the roots to enhance droplet capture.…”