Disposable Bioreactors for Plant Micropropagation and Mass Plant Cell Culture

Abstract

Different types of bioreactors are used at Nestlé R&D Centre - Tours for mass propagation of selected plant varieties by somatic embryogenesis and for large scale culture of plants cells to produce metabolites or recombinant proteins. Recent studies have been directed to cut down the production costs of these two processes by developing disposable cell culture systems. Vegetative propagation of elite plant varieties is achieved through somatic embryogenesis in liquid medium. A pilot scale process has recently been set up for the industrial propagation of Coffea canephora (Robusta coffee). The current production capacity is 3.0 million embryos per year. The pre-germination of the embryos was previously conducted by temporary immersion in liquid medium in 10-L glass bioreactors. An improved process has been developed using a 10-L disposable bioreactor consisting of a bag containing a rigid plastic box ('Box-in-Bag' bioreactor), insuring, amongst other advantages, a higher light transmittance to the biomass due to its horizontal design. For large scale cell culture, two novel flexible plastic-based disposable bioreactors have been developed from 10 to 100 L working volumes, validated with several plant species ('Wave and Undertow' and 'Slug Bubble' bioreactors). The advantages and the limits of these new types of bioreactor are discussed, based mainly on our own experience on coffee somatic embryogenesis and mass cell culture of soya and tobacco.

Perspectivas futuras e impacto social de las biotecnologías vegetales

“…En ese país, una parte significativa de la semilla certificada de caña de azúcar, plátanos, papa y otras especies llegó a producirse por vías biotecnológicas en las décadas de 1980-1990. En años recientes se han obtenido avances en el mundo en la propagación de numerosas especies, tales como diversos tipos de hortalizas (Estopá, 2005), plantas medicinales como Stemona tuberosa Lour (Biswas, Bari, Roy & Bhadra, 2011) Los esfuerzos actuales en la investigación se están dedicando a poner a punto tecnologías más complejas, que buscan reducir aún más el espacio dedicado a la multiplicación masiva, mediante el empleo de sistemas de inmersión temporal y biorreactores que se basan en el aprovechamiento de la embriogénesis somática (Ducos, Terrier & Courtois, 2009). Estos equipos tienen además como ventajas la posibilidad de propagar las plantas en escalas aún mayores que la micropropagación convencional, y de servir como biorreactores para obtener metabolitos útiles a las industrias química, alimentaria y farmacéutica.…”

Somatic Embryogenesis in Temporary Immersion Bioreactors

“…The TIB in the first group are the twin-flask bioreactor (Escalona et al 1999) and RALM bioreactor (Biorreactores RALM, Ralm Industria e Comércio ltda., Braz, http://ralm.nuvemshop.com.br/laboratorios-e-biofabricas/ biorreator-de-imersao-temporaria-ref-bio-001-2010/). The second group consists of the ebb-and-flow bioreactor (Ducos et al 2007b), RITA bioreactor , thermo-photo-bioreactor (Zuñiga Navarro et al 2013), bioreactor of immersion by bubbles (Scheidt et al 2009), SETIS (Vervit, Belgium, distributed by Duchefa Biochemie, The Netherlands), PLANTIMA bioreactor (A-Tech BioscientificCo., Ltd., Taiwan), PLANTFORM bioreactor (Plant Form AB, Sweden and TC propagation Ltd., Ireland), and the Box-in-Bag bioreactor (Ducos et al 2010). In the third group are the Rocker systems (Harris and Mason 1983), rotating drum bioreactor (Akita and Ohta), WAVE bioreactor (Eibl and Eibl), and BioMINT bioreactor (Robert et al 2006).…”

“…In C. arabica, using the RITA bioreactor, the conversion time of embryos to plants was 4 weeks compared to 6 weeks in semi-solid medium (Gatica-Arias et al 2008). In C. canephora, TIB allowed 95 % maturation from the torpedo stage to the cotyledonary stage (Ducos et al 2007b(Ducos et al , 2010. In C. arabica, the direct sowing of germinated embryos resulted in a highly successful conversion of embryos to plants (Etienne-Barry et al 1999;Albarrán et al 2005).…”

“…In B. gasipaes, a better response was achieved using the RITA bioreactor than using the twin-flask bioreactor (modified and unmodified), both in the multiplication of the cluster of somatic embryos and in converting them (Heringer et al 2014). For the pre-germination of coffee embryos, bioreactors called Glass Jar (Ducos et al 2007a, b) and box-in-bag (Ducos et al 2010) have been used successfully. Ducos et al (2010) found that horizontal bioreactors (box-in-bag) may be more convenient than vertical bioreactors because they provide a greater surface per volume.…”

“…For the pre-germination of coffee embryos, bioreactors called Glass Jar (Ducos et al 2007a, b) and box-in-bag (Ducos et al 2010) have been used successfully. Ducos et al (2010) found that horizontal bioreactors (box-in-bag) may be more convenient than vertical bioreactors because they provide a greater surface per volume. Furthermore, produced plantlets can be directly passed from laboratory to the greenhouse.…”