Disposable rocking bioreactors facilitate scaling up animal and plant cell biomass propagation and developing specified bioprocesses like manufacturing vaccines or chimeric antigen receptor (CAR) T cells. Future contexts for these bioreactors include supporting regenerative medicine, recognising metabolic responses of biochemically or mechanically stressed cells, continuously performing\u00a0in vitro<\/em>\u00a0bioprocesses, or cell-free protein synthesis systems.<\/p>\n\n\n\n Disposable rocking bioreactors allow efficient bubble-free surface aeration of cultures via 1D, 2D, or 3D oscillatory pivoting of foil-based bag-like containers, as opposed to bubble aeration occurring in typical bioreactors (vessels made with stainless steel or glass) equipped with internal mechanical stirrers, and a sparger or tube diffuser [1.<\/a>]. Such bubble-free surface aeration of the culture medium performed in rocking devices ensures mild conditions for\u00a0in vitro<\/em>\u00a0proliferating biomass in an environment with reduced levels of hydrodynamic shear stress. The range of shear stresses achieved in the rocking bioreactors is close to orbitally shaken and pneumatic bioreactors but significantly lower than noted in stirred tanks. The limited shear stress is the effect of limiting coalescence and disintegration of gas bubbles in rocked liquid. Furthermore, desirable culture conditions in rocking bioreactors can be achieved at a wide range of the values of the Reynolds number defined specifically for waving liquid, for example,\u00a0Re<\/em>L<\/sub>\u00a0=<\/em>\u00a0(sin<\/em>(\u03b1<\/em>)\u03c9L\/\u03bc<\/em>) [2.<\/a>]. Depending on the rocking angle (\u03b1<\/em>), the rocking frequency (\u03c9<\/em>), and the length of the rocked container (L<\/em>), various bioprocesses can be efficiently carried out at low (e.g.,\u00a0Re<\/em>L<\/sub>\u00a0=<\/em>\u00a0170 at\u00a0\u03b1<\/em>\u00a0= 2\u00b0\u00a0\u03c9<\/em>\u00a0= 2 min-1<\/sup>) or high (e.g.,\u00a0Re<\/em>L<\/sub>\u00a0=<\/em>\u00a020 000 at\u00a0\u03b1<\/em>= 12\u00b0\u00a0\u03c9<\/em>\u00a0= 40 min-1<\/sup>) turbulence in the same rocking bag-like container, according to the particular application\u2019s requirements. Because of that, disposable rocking bioreactors are suitable for\u00a0in vitro<\/em>\u00a0bioprocesses developed under a sufficient mild aeration intensity at desirable values of the volumetric mass transfer coefficient (k<\/em>L<\/sub>a<\/em>) and involving fragile or shear-sensitive animal or plant cells, aggregates, tissues, organs, and hybrid biomass-biomaterial 3D structures [3.<\/a>]. Unfortunately, the surface aeration occurring in rocking bioreactors is rather insufficient for cultures of biomass exhibiting high oxygen demand, such as microorganisms. Other drawbacks of these bioreactors include their incomplete engineering characterisation, causing uncertainty for potential users; the inefficient and still limited collaboration between biologists\/biotechnologists and small-scale bioprocess engineers; and steadily increasing price of rocking bioreactors and upgraded disposable containers, which limits their applications only to high-priced products. Overall, disposable rocking bioreactors are particularly useful in animal and plant cell cultures and are less suitable for microbial cultures.<\/p>\n\n\n\n We present modern, updated applications and progressive perspectives on the applications of disposable rocking bioreactors. The proposed discussion is supported by the concise but extensive summary of previously published literature on the applications of various commercially available systems of disposable rocking bioreactors and their progressive usage prospects.<\/p>\n\n\n\n A wide range of available working volumes of disposable bag-like containers (from 0.1 to 500 dm3<\/sup>) encourage the practical applicability of disposable bioreactors for bioprocess implementation. In the market, different capacities of disposable containers are available, that is, 0.1\u20131 dm3<\/sup>, 0.5\u20135 dm3<\/sup>, 2\u201320 dm3<\/sup>, 5\u201350 dm3<\/sup>, 10\u2013100 dm3<\/sup>, 20\u2013200 dm3<\/sup>, and 50\u2013500 dm3<\/sup>, but most commonly used are bag-like containers with 0.1\u20131 dm3<\/sup>\u00a0and 5\u201350 dm3<\/sup>\u00a0capacities. Nowadays, in the case of animal cell cultures, the use of systems equipped with disposable bag-like containers is the most common way for the first step of bioreactor-based scaling up bioprocesses from the volume of millilitres, that is, from various multiwall plates, culture flasks, roller bottles, Erlenmeyer flasks (i.e., from ca. 0.1 dm3<\/sup>) into litre-scale cultures (i.e., up to 100 dm3<\/sup>). The other systems also applied to efficient scaling up animal cell cultures, that is, multiplied culture flasks or roller bottles, are less efficient and limited to a few litres of volume. Modern applications of devices supporting the oscillatory rocking of bag-like containers are reviewed in\u00a0Table 1<\/a>.<\/p>\n\n\n\n Open table in a new tab<\/a><\/p>\n\n\n\n The data presented in\u00a0Table 1<\/a>\u00a0is limited only to the most commonly used rocking bioreactors, two models of the 1D rocking system: BIOSTAT RM of Sartorius and\u00a0ReadyToProcess<\/em>\u00a0WAVE (formerly WAVE Bioreactor) of Cytiva and one model of the 2D rocking system: Cell-tainer of Cell-tainer Biotech BV. Only one model of 3D rocking bioreactors is available on the market (Allegro\u2122 XRS 25 produced by Pall); however, no published data supports its application in bioprocessing. The differences in the mechanisms of culture medium mixing in 1D, 2D, and 3D disposable rocking bioreactors are presented in\u00a0Box 1<\/a>.<\/p>\n\n\n\n Over the past 5 years, disposable rocking systems have been applied for scaling up bioprocesses incorporating various types of cells, for example, human [4.<\/a>,5.<\/a>], mammalian [3.<\/a>], insect [6.<\/a>], plant cells and organs [7.<\/a>,8.<\/a>], bacteria [9.<\/a>,10.<\/a>], and\u00a0Streptomyces<\/em>\u00a0[11.<\/a>], as well as for maintaining cell-free system containing purified lysates of plant cells [12.<\/a>]. The most recently published scientific reports or experimental protocols present valuable and precise quantitative data on highly specified bioprocesses developed in oscillatory rocked disposable bag-like containers, emphasising their suitability for the propagation of fragile biomass. Such applications go hand-in-hand with already extensively implemented or hypothetically expected demands of the biopharmaceutical industry, including high-cell-density batch and fed-batch cultures of shear-sensitive or structurally integrated forms of biomass, including the following:<\/p>\n\n\n\n The extensive use of disposable rocking bioreactors for the large-scale production of biotic agents for vaccines in the era of the recent COVID-19 pandemic emphatically confirmed their practical applicability in the biopharmaceutical industry. It unambiguously proved their flexibility for pharma applications in large-scale (even global!) bioproduct production according to methodologies requiring rapid implementation of new or nontypical techniques or devices. However, currently and probably in the future, the most intensively developed branch of rocking bioreactors\u2019 applicability may be the proliferation of CAR T cells in anticancer therapies. New generations of customer-tailored bag-like containers will also be required in this case.<\/p>\n\n\n\n As for future applications, we hypothesise that in addition to the existing ones listed previously, rocking bioreactors will efficiently support regenerative medicine in the scope of enhanced colonisation of biomaterial-based implants with cells isolated from the patients to reconstruct tissue losses, such as bones, cartilage, muscles, or blood vessels. Rocking devices supported with custom-tailored disposable containers are recognised as an invaluable tool in scaling up cultures of human stem cells (pluripotent and adult ones), their further propagation and controlled differentiation, performed not only on a laboratory scale but also for commercial manufacturing of allogenic pluripotent stem cell-based products. In the case of the biopharmaceutical industry, atypical mixing performance resulting from the waving liquid agitated in disposable bag-like containers has already been identified as the physical causative agent forcing the metabolic response of biomass (i.e., prokaryotic and eukaryotic cells) to shear stress [11.<\/a>]. This promising research direction should be developed in research and applications shortly. Simplified handling of disposable bag-like containers makes rocking bioreactors crucial elements of the single-use strategies for continuous bioprocesses applied in, for example, monoclonal antibodies production [13.<\/a>],\u00a0in situ<\/em>\u00a0biomass elicitation and separation of bioactive compounds of botanical origin [7.<\/a>], separation of metabolites from culture systems [11.<\/a>], and in cell-free systems for protein biosynthesis in purified lysates of prokaryotic or eukaryotic cells [12.<\/a>]. Moreover, appropriate sterile usage of disposable bag-like containers lets multiple uses of them as closed plug-and-play oscillatory driven mixers for upstream or downstream processing, reducing their unit costs and increasing the development of intensified bioprocesses in recent years.<\/p>\n\n\n\n We also recommend computational fluid dynamics (CFD)-supported designing and prototyping of new generations of shape-modified disposable bag-like containers manufactured with foils containing structural 3D elements, which passively improve mixing performance and aeration efficiency of liquids. Developing such modified containers will widen the application of rocking bioreactors for bioprocesses with biomass characterised by high oxygen demand (i.e., microorganisms) and fragile biomass cultured at high cell density (i.e., callus cells or protoplasts, animal or insect cells).<\/p>\n\n\n\n Disposable rocking bioreactors are widely used in developing and scaling up in vitro<\/em> bioprocesses. Until recently, such devices equipped with single-use bag-like containers were mainly applied to propagate fragile forms of biomass. During the COVID-19 pandemic, virus propagation in isolated mammalian cells and mRNA strand production for anti-SARS-CoV-2 vaccine manufacturing was the leading global-scale application of disposable bioreactors. Recent publications signal a likely direction for widening the applicability of disposable rocking devices in supporting regenerative medicine and strategies for continuous bioprocessing and developing systems for metabolic responses of non-natively stress-induced cells maintained in vitro<\/em>. Nevertheless, in all possible perspectives for disposable rocking bioreactors, the knowledge of the mass transfer characteristics and mixing performance in applied bag-like containers will be crucial.<\/p>\n","protected":false},"excerpt":{"rendered":" Abstract Disposable rocking bioreactors facilitate scaling up animal and plant cell biomass propagation and developing specified bioprocesses like manufacturing vaccines or chimeric antigen receptor (CAR) T cells. Future contexts for these bioreactors include supporting regenerative medicine, recognising metabolic responses of biochemically or mechanically stressed cells, continuously performing\u00a0in vitro\u00a0bioprocesses, or cell-free protein synthesis systems. Disposable rocking […]<\/p>\n","protected":false},"author":3,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_seopress_robots_primary_cat":"none","nova_meta_subtitle":"Recent applications and progressive perspectives","footnotes":""},"categories":[5572],"tags":[16380,11865,5842,22839,12420],"supplier":[],"class_list":["post-132804","post","type-post","status-publish","format-standard","hentry","category-bio-based","tag-biobased","tag-biochemistry","tag-biomass","tag-bioprocessing","tag-bioreactor"],"_links":{"self":[{"href":"https:\/\/renewable-carbon.eu\/news\/wp-json\/wp\/v2\/posts\/132804","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/renewable-carbon.eu\/news\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/renewable-carbon.eu\/news\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/renewable-carbon.eu\/news\/wp-json\/wp\/v2\/users\/3"}],"replies":[{"embeddable":true,"href":"https:\/\/renewable-carbon.eu\/news\/wp-json\/wp\/v2\/comments?post=132804"}],"version-history":[{"count":0,"href":"https:\/\/renewable-carbon.eu\/news\/wp-json\/wp\/v2\/posts\/132804\/revisions"}],"wp:attachment":[{"href":"https:\/\/renewable-carbon.eu\/news\/wp-json\/wp\/v2\/media?parent=132804"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/renewable-carbon.eu\/news\/wp-json\/wp\/v2\/categories?post=132804"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/renewable-carbon.eu\/news\/wp-json\/wp\/v2\/tags?post=132804"},{"taxonomy":"supplier","embeddable":true,"href":"https:\/\/renewable-carbon.eu\/news\/wp-json\/wp\/v2\/supplier?post=132804"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}Disposable rocking bioreactors<\/h2>\n\n\n\n
Modern updated applications of disposable rocking bioreactors<\/h3>\n\n\n\n
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Future perspectives for disposable rocking systems<\/h3>\n\n\n\n
Concluding remarks<\/h3>\n\n\n\n