Suspension cell culture in microgravity and development of a space bioreactor

NASA has methodically developed unique suspension type cell and recovery apparatus culture systems for bioprocess technology experiments and production of biological products in microgravity. The first space bioreactor has been designed for microprocessor control, no gaseous headspace, circulation and resupply of culture medium, and slow mixing in very low shear regimes. Various ground based bioreactors are being used to test reactor vessel design, on-line sensors, effects of shear, nutrient supply, and waste removal from continuous culture of human cells attached to microcarriers. The small (500 ml) bioreactor is being constructed for flight experiments in the Shuttle middeck to verify systems operation under microgravity conditions and to measure the efficiencies of mass transport, gas transfer, oxygen consumption, and control of low shear stress on cells

Astex microgravity experiment: simulated asteroid regoliths

AstEx is a microgravity experiment selected to fly on ESA's 51st Microgravity Research Campaign in November 2009. The experiment will investigate the dynamics of regolith on asteroid surfaces. Despite their very low surface gravities, asteroids exhibit a number of different geological processes involving granular matter. Understanding the mechanical response of this granular material subject to external forces in microgravity conditions is vital to the design of a successful asteroid sub-surface sampling mechanism, and in the interpretation of the fascinating geology on an asteroid. The AstEx experiment uses a microgravity modified Taylor-Couette shear cell to investigate granular flow caused by shear forces under the conditions of parabolic flight microgravity. It is intended to determine how a steady state granular flow is achieved in microgravity conditions, and what effect prior shear history has on the timescales involved in initiating a steady state flow in a granular material. Presented are the technical details of the AstEx experimental design with particular emphasis on how the team have designed the equipment specifically for the parabolic flight microgravity environment

A Simulated Microgravity Environment Causes a Sustained Defect in Epithelial Barrier Function.

Intestinal epithelial cell (IEC) junctions constitute a robust barrier to invasion by viruses, bacteria and exposure to ingested agents. Previous studies showed that microgravity compromises the human immune system and increases enteropathogen virulence. However, the effects of microgravity on epithelial barrier function are poorly understood. The aims of this study were to identify if simulated microgravity alters intestinal epithelial barrier function (permeability), and susceptibility to barrier-disrupting agents. IECs (HT-29.cl19a) were cultured on microcarrier beads in simulated microgravity using a rotating wall vessel (RWV) for 18 days prior to seeding on semipermeable supports to measure ion flux (transepithelial electrical resistance (TER)) and FITC-dextran (FD4) permeability over 14 days. RWV cells showed delayed apical junction localization of the tight junction proteins, occludin and ZO-1. The alcohol metabolite, acetaldehyde, significantly decreased TER and reduced junctional ZO-1 localization, while increasing FD4 permeability in RWV cells compared with static, motion and flask control cells. In conclusion, simulated microgravity induced an underlying and sustained susceptibility to epithelial barrier disruption upon removal from the microgravity environment. This has implications for gastrointestinal homeostasis of astronauts in space, as well as their capability to withstand the effects of agents that compromise intestinal epithelial barrier function following return to Earth

Design challenges for space bioreactors

The design of bioreactors for operation under conditions of microgravity presents problems and challenges. Absence of a significant body force such as gravity can have profound consequences for interfacial phenomena. Marangoni convection can no longer be overlooked. Many speculations on the advantages and benefits of microgravity can be found in the literature. Initial bioreactor research considerations for space applications had little regard for the suitability of the designs for conditions of microgravity. Bioreactors can be classified in terms of their function and type of operation. The complex interaction of parameters leading to optimal design and operation of a bioreactor is illustrated by the JSC mammalian cell culture system. The design of a bioreactor is strongly dependent upon its intended use as a production unit for cell mass and/or biologicals or as a research reactor for the study of cell growth and function. Therefore a variety of bioreactor configurations are presented in rapid summary. Following this, a rationale is presented for not attempting to derive key design parameters such as the oxygen transfer coefficient from ground-based data. A set of themes/objectives for flight experiments to develop the expertise for design of space bioreactors is then proposed for discussion. These experiments, carried out systematically, will provide a database from which engineering tools for space bioreactor design will be derived

Modelling the Interfacial Flow of Two Immiscible Liquids in Mixing Processes

This paper presents an interface tracking method for modelling the flow of immiscible metallic liquids in mixing processes. The methodology can provide an insight into mixing processes for studying the fundamental morphology development mechanisms for immiscible interfaces. The volume-of-fluid (VOF) method is adopted in the present study, following a review of various modelling approaches for immiscible fluid systems. The VOF method employed here utilises the piecewise linear for interface construction scheme as well as the continuum surface force algorithm for surface force modelling. A model coupling numerical and experimental data is established. The main flow features in the mixing process are investigated. It is observed that the mixing of immiscible metallic liquids is strongly influenced by the viscosity of the system, shear forces and turbulence. The numerical results show good qualitative agreement with experimental results, and are useful for optimisating the design of mixing casting processes

Monitoring Three-Dimensional Packings in Microgravity

We present results from experiments with granular packings in three dimensions in microgravity as realized on parabolic flights. Two different techniques are employed to monitor the inside of the packings during compaction: (1) X-ray radiography is used to measure in transmission the integrated fluctuations of particle positions. (2) Stress-birefringence in three dimensions is applied to visualize the stresses inside the packing. The particle motions below the transition into an arrested packing are found to produce a well agitated state. At the transition, the particles lose their energy quite rapidly and form a stress network. With both methods, non-arrested particles (rattlers) can be identified. In particular, it is found that rattlers inside the arrested packing can be excited to appreciable dynamics by the rest-accelerations (g-jitter) during a parabolic flight without destroying the packings. At low rates of compaction, a regime of slow granular cooling is identified. The slow cooling extends over several seconds, is described well by a linear law, and terminates in a rapid final collapse of dynamics before complete arrest of the packing.Comment: 8 pages, 8 figure

Design concepts for bioreactors in space

Microbial food sources are becoming viable and more efficient alternatives to conventional food sources especially in the context of Closed Ecological Life Support Systems (CELSS) in space habitats. Since bioreactor designs for terrestrial operation will not readily apply to conditions of microgravity, there is an urgent need to learn about the differences. These differences cannot be easily estimated due to the complex nature of the mass transport and mixing mechanisms in fermenters. Therefore, a systematic and expeditious experimental program must be undertaken to obtain the engineering data necessary to lay down the foundations of designing bioreactors for microgravity. Two bioreactor design concepts presented represent two dissimilar approaches to grappling with the absence of gravity in space habitats and deserve to be tested for adoption as important components of the life support function aboard spacecrafts, space stations and other extra-terrestrial habitats

Analysis of the characteristic lengths in the bubble and slug flow regimes generated in a capillary T-junction

We present an analysis of the geometry of the continuous and disperse phases in the bubble and slug flow regimes in air–water mixtures generated in a capillary T-junction of 1 mm internal diameter. Bubble size dispersion is very low in the considered flow patterns. The concept of unit cell is used to identify two characteristic lengths of the two-phase flow, namely, the unit cell length and the bubble length. The relationship between these lengths and the gas and liquid superficial velocities, gas mean velocity, bubble generation frequency and volume average void fraction is analysed. We conclude that in the considered configuration the unit cell and bubble lengths can be predicted either by the ratio of the gas–liquid superficial velocities or the volume average void fraction.Peer ReviewedPreprin

Media Optimization, Strain Compatibility, and Low-Shear Modeled Microgravity Exposure of Synthetic Microbial Communities for Urine Nitrification in Regenerative Life-Support Systems

Urine is a major waste product of human metabolism and contains essential macro- and micronutrients to produce edible microorganisms and crops. Its biological conversion into a stable form can be obtained through urea hydrolysis, subsequent nitrification, and organics removal, to recover a nitrate-enriched stream, free of oxygen demand. In this study, the utilization of a microbial community for urine nitrification was optimized with the focus for space application. To assess the role of selected parameters that can impact ureolysis in urine, the activity of six ureolytic heterotrophs (Acidovorax delafieldii, Comamonas testosteroni, Cupriavidus necator, Delftia acidovorans, Pseudomonas fluorescens, and Vibrio campbellii) was tested at different salinities, urea, and amino acid concentrations. The interaction of the ureolytic heterotrophs with a nitrifying consortium (Nitrosomonas europaea ATCC 19718 and Nitrobacter winogradskyi ATCC 25931) was also tested. Lastly, microgravity was simulated in a clinostat utilizing hardware for in-flight experiments with active microbial cultures. The results indicate salt inhibition of the ureolysis at 30 mS cm(-1), while amino acid nitrogen inhibits ureolysis in a strain-dependent manner. The combination of the nitrifiers with C. necator and V. campbellii resulted in a complete halt of the urea hydrolysis process, while in the case of A. delafieldii incomplete nitrification was observed, and nitrite was not oxidized further to nitrate. Nitrate production was confirmed in all the other communities; however, the other heterotrophic strains most likely induced oxygen competition in the test setup, and nitrite accumulation was observed. Samples exposed to low-shear modeled microgravity through clinorotation behaved similarly to the static controls. Overall, nitrate production from urea was successfully demonstrated with synthetic microbial communities under terrestrial and simulated space gravity conditions, corroborating the application of this process in space

Mathematical modeling of the flow field and particle motion in a rotating bioreactor at unit gravity and microgravity

The biotechnology group at NASA Johnson Space Center is developing systems for culturing mammalian cells that stimulate some aspect of microgravity and provide a low shear environment for microgravity-based studies on suspension and anchorage dependent cells. The design of these vessels for culturing cells is based on the need to suspend cells and aggregates of cells and microcarrier beads continually in the culturing medium. The design must also provide sufficient circulation for adequate mass transfer of nutrients to the cells and minimize the total force on the cells. Forces, resulting from sources such as hydrodynamic fluid shear and collisions of cells and walls of the vessels, may damage delicate cells and degrade the formation of three dimensional structures. This study examines one particular design in both unit gravity and microgravity based on two concentric cylinders rotating in the same direction at different speeds to create a Couette flow between them. A numerical simulation for the flow field and the trajectories of particles in the vessel. The flow field for the circulation of the culturing medium is modeled by the Navier-Stokes equations. The forces on a particle are assumed to be drag from the fluid's circulation, buoyancy from the gravitational force and centrifugal force from the rotation of the vessel. The problem requires first solving the system of partial differential equations for the fluid flow by a finite difference method and then solving the system of ordinary differential equations for the trajectories by Gear's stiff method. Results of the study indicate that the trajectories in unit gravity and microgravity are very similar except for small spatial deviations on the fast time scale in unit gravity. The total force per unit cross sectional area on a particle in microgravity, however, is significantly smaller than the corresponding value in unit gravity, which is also smaller than anticipated. Hence, this study indicates that this design for a bioreactor with optimal rates of rotation can provide a good environment for culturing cells in microgravity with adequate circulation and minimal force on the cells