Large-scale Production Systems Research Articles

Cost-effective Purification of Membrane Proteins using a Dual-detergent Strategy.

Understanding the mechanisms of membrane protein function is critical for biomedical research and drug discovery as membrane proteins constitute ∼30% of the proteins encoded by the genomes of both lower and higher organisms and are targets for two-thirds of approved drugs worldwide. Significant progress has been made in engineering host expression systems for large-scale production of membrane proteins and in determining their three-dimensional high-resolution structures. Despite these efforts, the study of membrane proteins at the atomic level is challenging due to poor expression and extraction, low yields of functional protein, and the complexity and heterogeneity of source membranes. Structural and spectroscopic studies of any membrane protein require that the protein be extracted from its native membranes into a membrane-mimetic stable environment, which is often achieved by the use of detergents. Unfortunately, there is no magic detergent that can extract all membrane proteins and successful extraction often requires a thorough screen of detergents. Furthermore, membrane protein purification in general and the detergents used are very expensive, which puts a financial constraint on sophisticated membrane protein studies. To overcome this hurdle, a dual-detergent strategy has recently been developed and successfully applied to purify various classes of pure, stable, and functionally relevant membrane proteins in a cost-effective manner. This strategy uses an inexpensive detergent for solubilization of the desired protein from membranes and a second detergent during protein purification. In the Basic Protocol, we describe the dual-detergent strategy to significantly reduce the overall purification cost of a bacterial membrane protein using the magnesium ion channel MgtE as an example. Support Protocols are also provided for selecting a suitable E. coli strain for protein expression and the optimal detergent(s) for membrane protein solubilization. © 2022 Wiley Periodicals LLC. Basic Protocol: Expression, membrane solubilization, and cost-effective purification of MgtE Support Protocol 1: Selecting a suitable E. coli strain for optimal protein expression Support Protocol 2: Identification of suitable detergents for membrane protein solubilization.

Dextran sulfate prevents excess aggregation of human pluripotent stem cells in 3D culture by inhibiting ICAM1 expression coupled with down-regulating E-cadherin through activating the Wnt signaling pathway

BackgroundHuman pluripotent stem cells (hPSCs) have great potential in applications for regenerative medicine and drug development. However, 3D suspension culture systems for clinical-grade hPSC large-scale production have been a major challenge. Accumulating evidence has demonstrated that the addition of dextran sulfate (DS) could prevent excessive adhesion of hPSCs from forming larger aggregates in 3D suspension culture. However, the signaling and molecular mechanisms underlying this phenomenon remain elusive.MethodsBy using a cell aggregate culture assay and separating big and small aggregates in suspension culture systems, the potential mechanism and downstream target genes of DS were investigated by mRNA sequence analysis, qRT-PCR validation, colony formation assay, and interference assay.ResultsSince cellular adhesion molecules (CAMs) play important roles in hPSC adhesion and aggregation, we assumed that DS might prevent excess adhesion through affecting the expression of CAMs in hPSCs. As expected, after DS treatment, we found that the expression of CAMs was significantly down-regulated, especially E-cadherin (E-cad) and intercellular adhesion molecule 1 (ICAM1), two highly expressed CAMs in hPSCs. The role of E-cad in the adhesion of hPSCs has been widely investigated, but the function of ICAM1 in hPSCs is hardly understood. In the present study, we demonstrated that ICAM1 exhibited the capacity to promote the adhesion in hPSCs, and this adhesion was suppressed by the treatment with DS. Furthermore, transcriptomic analysis of RNA-seq revealed that DS treatment up-regulated genes related to Wnt signaling resulting in the activation of Wnt signaling in which SLUG, TWIST, and MMP3/7 were highly expressed, and further inhibited the expression of E-cad.ConclusionOur results demonstrated that DS played an important role in controlling the size of hPSC aggregates in 3D suspension culture by inhibiting the expression of ICAM1 coupled with the down-regulation of E-cad through the activation of the Wnt signaling pathway. These results represent a significant step toward developing the expansion of hPSCs under 3D suspension condition in large-scale cultures.

Fertilizer type and humic acid improve the growth responses, nutrient uptake, and essential oil content on Coriandrum sativum L.

In recent decades, the over-use of chemical fertilizers has imposed many environmental challenges worldwide. Nowadays, organic fertilizers such as vermicompost and livestock manure have gained a huge interest in sustainable agricultural systems. A 2-year field research was conducted as factorial based on a randomized complete block design to assay the fertilizer and humic acid (HA) efficiency on the growth responses and essential oil composition of Coriandrum sativum. The treatments were different fertilizer sources (livestock manure, vermicompost, and chemical fertilizers) and humic acid fertigation before and at the beginning of the flowering stage. The highest protein content was observed under vermicompost × HA application before flowering (0.118 μmol L−1 and 0.128 μmol L−1, respectively). Moreover, the co-application of organic fertilizers × HA at the beginning of flowering resulted in a significant increase in the photosynthetic pigments and N, P, K, Fe, Zn, and Mn content. According to the GC-FID and GC–MS analysis, linalool (55.91–63.19%), γ-terpinene (4.65–6.13%), α-pinene (2.64–5.74%), geranyl acetate (3.49–5.51%), 2-dodecanal (2.92–4.46%), menthol (1.33–3.90%), p-cymene (1.73–2.24%), and geraniol (1.25–2.15%) were the main essential oil constituents. The top linalool content was obtained by using chemical fertilizers and vermicompost × HA at the flowering onset stage. In general, the results revealed that chemical fertilizers could be replaced with vermicompost × HA and their co-application positively influenced the growth responses and the essential oil composition of coriander. Furthermore, the results obtained would be advisable to the extension section and the pioneer farmers to amend the large-scale production systems in favor of environmental health.

Biomass-derived carbons physically activated in one or two steps for CH4/CO2 separation

The present study aims at evaluating the suitability of producing activated carbons (ACs) derived from wheat straw by a one-step synthesis approach, as an alternative to more conventional two steps production processes (i.e., pyrolysis and subsequent activation). The performance of the produced ACs, in one or two steps, as sustainable and selective CO2 adsorbents for CH4/CO2 separation is compared. In addition, the influence of pyrolysis conditions on the properties of the resulting two-step ACs is carefully analyzed. We show that the biochar-based precursors of ACs presenting the best textural properties were obtained under mild conditions of maximum temperature and absolute pressure during pyrolysis. The one-step ACs were fully comparable —in terms of textural properties as well as CO2 uptake and selectivity— to those produced by the more conventional two-step synthesis process. In addition, results obtained from breakthrough curve simulations highlight that the best AC in terms of CH4 recovery under dynamic conditions was produced by a one-step activation. Therefore, the one-step process appears to be as an attractive route for the production of engineered carbon materials, which can lead to significant cost savings in large-scale production systems.

Potential Use of Quartzipisamment under Agroforestry and Silvopastoral System for Large-Scale Production in Brazil

The need to put into practice sustainable agricultural production systems has been supported by agroecology science that seeks to optimize land use to food production with the lowest impact on soil. This study evaluated soil quality, based on physical and chemical attributes, in agroforestry (AGF) and silvopastoral (SILVP) systems developed for large-scale food production. The study was carried out in the municipality of Itirapina, state of São Paulo, in two areas with AGF and SILVP system, compared to an area with a forest fragment and another with pasture in a Quartzipisamment Sand Neosol. The soil collections were carried out in the layers of 0.00–0.05, 0.05–0.10, 0.10–0.20, and 0.20–0.40 m, where physical soil attributes were evaluated (total porosity, microporosity, and microporosity, density, mean diameter of aggregates) as well as chemical attributes (macro- and micronutrients), in addition to carbon and nitrogen storage. To interpret the data, Tukey’s test was applied to compare means, and principal component analysis was used to better characterize the study environments. The results showed that agroforestry and silvopastoral systems developed for large-scale production are efficient in improving chemical and physical attributes that reflect on soil quality, especially in the superficial layers of the soil, overcoming pasture and the natural regeneration process. Carbon and nitrogen storage were the main variables that differentiated the production systems, highlighting the importance of the AGF and SILVP systems as more sustainable agricultural intensification strategies, even in soils of low agricultural suitability.

Efficient production of inositol from glucose via a tri-enzymatic cascade pathway

Inositol is an essential intermediate in cosmetics, food, medicine and other industries. However, developing an efficient biotransformation system for large-scale production of inositol remains challenging. Herein, a tri-enzymatic cascade route with three novel enzymes including polyphosphate glucokinase (PPGK) from Thermobifida fusca, inositol 3-phosphate synthase (IPS) from Archaeoglobus profundus DSM 5631 and inositol monophosphatase (IMP) from Thermotoga petrophila RKU-1 was designed and reconstructed for the production of inositol from glucose. The problem of poor cooperativity of the cascade reactions was addressed by ribosome binding site (RBS) optimization of PPGK and replication of IPS. Under the optimum biotransformation conditions, the engineered whole-cell immobilized with colloidal chitin transformed 120 g/L glucose to 110.8 g/L inositol with 92.3% conversion in four cycles of reuse, representing the highest titer of inositol to date. Furthermore, this is the first study for inositol production using a three-enzyme coordinated immobilized single-cell.

Digital Twin-Driven Performance Optimization for Hazardous Waste Landfill Systems

Hazardous waste landfill is the final way to dispose of hazardous waste and its environmental risk is different from that of domestic waste landfill. Its dangerous characteristics will not decrease with time, but will exist for a long. The environmental risks of hazardous waste landfills are concentrated in the probability risk of leakage of the impermeable layer. Therefore, it is of great significance to calculate the environmental risk of the leakage source intensity of typical hazardous waste landfills. In order to adapt to the personalized customer needs caused by social and economic development and to pursue new competitive advantages, the production mode of manufacturing enterprises is gradually changing to a small-batch and multi-variety customized mode. The complex and dynamic production environment caused by determinism puts forward higher requirements for the operation control accuracy of large-scale multiunit production systems. To support the efficient and coordinated operation of each link is a key challenge for process control of large-scale multiunit production logistics systems. This article proposes to accelerate the integration of information technology and manufacturing technology to promote the application of intelligent manufacturing process, intelligent logistics management, and other technologies in the production process. The research object is to study the multiunit linkage optimization problem under the framework of digital twin technology, through the real-time collaborative decision-making and feedback execution of the production, transportation, and warehousing multi-decision links to solve the “production-transportation-storage” decision-making in a dynamic environment. We analyzed the problems brought about by the dynamic interference of the execution process on the “production-transportation-storage” linkage operation process of the industrial park and analyzed the challenges of realizing the online linkage decision-making of “production-transportation-storage.” Based on the classic digital twin framework, a cross-unit digital twin linkage decision information architecture is proposed. Compared with the conclusions of the original environmental impact assessment report of the project, the post-environmental impact assessment results basically conform to the relevant descriptions, and a few nonconformities have been resolved during the project operation. The environmental protection measures of this project are relatively complete, which can meet various environmental protection requirements and enterprise development needs during the operation period of the project. For some environmental risks found in the process of project operation, evasion measures are proposed to reduce the possibility of environmental pollution accidents, and emergency measures can be taken in time when accidents occur. The practice of post-assessment of the environmental impact of this hazardous waste landfill is generally successful, and it has certain reference significance for the subsequent post-assessment of the environmental impact of other hazardous waste landfills.

Recombinant Spider Silk: Promises and Bottlenecks.

Spider silk threads have exceptional mechanical properties such as toughness, elasticity and low density, which reach maximum values compared to other fibre materials. They are superior even compared to Kevlar and steel. These extraordinary properties stem from long length and specific protein structures. Spider silk proteins can consist of more than 20,000 amino acids. Polypeptide stretches account for more than 90% of the whole protein, and these domains can be repeated more than a hundred times. Each repeat unit has a specific function resulting in the final properties of the silk. These properties make them attractive for innovative material development for medical or technical products as well as cosmetics. However, with livestock breeding of spiders it is not possible to reach high volumes of silk due to the cannibalistic behaviour of these animals. In order to obtain spider silk proteins (spidroins) on a large scale, recombinant production is attempted in various expression systems such as plants, bacteria, yeasts, insects, silkworms, mammalian cells and animals. For viable large-scale production, cost-effective and efficient production systems are needed. This review describes the different types of spider silk, their proteins and structures and discusses the production of these difficult-to-express proteins in different host organisms with an emphasis on plant systems.

Improved utilization of IrOx on Ti4O7 supports in membrane electrode assembly for polymer electrolyte membrane water electrolyzer

Recently, since the continuous rise in cost and scarcity of iridium, it is necessary to minimize the iridium-loading in the membrane electrode assembly (MEA) for large-scale green hydrogen production systems. In this study, IrOx/Ti4O7 catalysts were prepared to increase the utilization of IrOx in the MEA by evenly dispersing the structurally layered IrOx on Ti4O7 support via a modified simple solution-reduction method. Among the catalysts with various iridium/support ratios, the IrOx/Ti4O7(7:3) catalyst demonstrated the optimal balance of catalytic activity, catalytic stability, and corrosion resistance. IrOx/Ti4O7(7:3) catalyst demonstrated a higher mass activity of 372 mA mg-1 at 1.55 V, stable durability period of > 12 h under accelerated stability test (AST), and a low iridium dissolution rate of 0.078 ppbIr h-1. A single-cell test was further conducted using the IrOx/Ti4O7(7:3) catalyst to reveal its catalytic activity in the MEA. Consequently, the mass activity of the IrOx/Ti4O7(7:3) catalyst was found to increase by 139% in comparison to a black IrOx/None catalyst. Additionally, in the single-cell durability test (applied 1 A cm-2 for 50 h), the IrOx/Ti4O7(7:3) catalyst demonstrated a degradation rate of only 0.6 mV h-1 at 1000 A g-1 which can be estimated to have reasonable stability. Therefore, it is anticipated that the IrOx/Ti4O7(7:3) catalyst will reduce the amount of high-cost iridium in the MEA and will result in a low-cost and efficient polymer electrolyte membrane water electrolyzer system for large-scale hydrogen production.

Large-scale hydrogen production via water electrolysis: a techno-economic and environmental assessment

This work quantifies current and future costs as well as environmental burdens of large-scale hydrogen production systems on geographical islands, which exhibit high renewable energy potentials and could act as hydrogen export hubs.

Technoeconomics of large-scale clean hydrogen production – A review

Hydrogen is widely used in many industries, yet its role in the clean energy transition goes beyond being an element of these industries. Near-term practical large-scale clean hydrogen production can be made available by involving nuclear, solar, and other renewable energy sources in the process of hydrogen production, and coupling their energy systems to sustainable carbon-free hydrogen technologies. This requires further investigation and assessment of the different alternatives to achieve clean hydrogen using these pathways. This paper assesses the technoeconomics of promising hydrogen technologies that can be coupled to nuclear and solar energy systems for large-scale hydrogen production. It also provides an overview of the design, status and advances of these technologies.

A stacked deep learning approach to cyber-attacks detection in industrial systems: application to power system and gas pipeline systems.

Presently, Supervisory Control and Data Acquisition (SCADA) systems are broadly adopted in remote monitoring large-scale production systems and modern power grids. However, SCADA systems are continuously exposed to various heterogeneous cyberattacks, making the detection task using the conventional intrusion detection systems (IDSs) very challenging. Furthermore, conventional security solutions, such as firewalls, and antivirus software, are not appropriate for fully protecting SCADA systems because they have distinct specifications. Thus, accurately detecting cyber-attacks in critical SCADA systems is undoubtedly indispensable to enhance their resilience, ensure safe operations, and avoid costly maintenance. The overarching goal of this paper is to detect malicious intrusions that already detoured traditional IDS and firewalls. In this paper, a stacked deep learning method is introduced to identify malicious attacks targeting SCADA systems. Specifically, we investigate the feasibility of a deep learning approach for intrusion detection in SCADA systems. Real data sets from two laboratory-scale SCADA systems, a two-line three-bus power transmission system and a gas pipeline are used to evaluate the proposed method’s performance. The results of this investigation show the satisfying detection performance of the proposed stacked deep learning approach. This study also showed that the proposed approach outperformed the standalone deep learning models and the state-of-the-art algorithms, including Nearest neighbor, Random forests, Naive Bayes, Adaboost, Support Vector Machine, and oneR. Besides detecting the malicious attacks, we also investigate the feature importance of the cyber-attacks detection process using the Random Forest procedure, which helps design more parsimonious models.

Unravelling the anaerobic digestion ‘black box’: Biotechnological approaches for process optimization

Increasing global demand for green energy has necessitated more research interest in anaerobic digestion. Due to the dual benefits of anaerobic digestion, in organic waste management and energy generation, it is rated among the best renewable energy generation options. An important challenge that limits acceptability and adoption of this technology is the quality and quantity of anaerobic digestion products, especially biogas and digestate. A practical solution to this challenge is process optimization. However, process optimization is not easily achieved due to the complexity of the microbiome driving the anaerobic digestion process, which has contributed to anaerobic digesters being considered ‘black boxes’. Unravelling the ‘black box’ would aid in fine-tuning operational conditions, process parameters, intra- and inter-phase interactions as well as rate limiting factors during each phase of the production cycle, thereby optimizing the anaerobic digestion process. To achieve this level of technological advancement, cooperation between experts that understand both the microbial processes and technological design of digesters is important. When factors are optimized, implementation of the optimized conditions through automation may prevent process errors and failures, thereby enhancing quality and quantity of anaerobic digestion products. In addition, process optimization should also reflect economic viability vis-à-vis small-scale and large-scale biogas production systems. This review provides a detailed overview of individual factors that could affect the performance of the anaerobic digestion process with critical interpretations and suggestions. This work could be employed as a good base to build integrative models or ecological indices that interrelate these factors.

Integration of Algal Biofuels With Bioremediation Coupled Industrial Commodities Towards Cost-Effectiveness

Microalgae offer a great potential to contribute significantly as renewable fuels and documented as a promising platform for algae-based bio refineries. They provide solutions to mitigate the environmental concerns posed by conventional fuel sources; however, the production of microalgal biofuels in large scale production system encounters few technical challenges. High quantity of nutrients requirements and water cost constrain the scaling up microalgal biomass to large scale commercial production. Crop protection against biomass losses due to grazers or pathogens is another stumbling block in microalgal field cultivation. With our existing technologies, unless coupled with high-value or mid-value products, algal biofuel cannot reach the economic target. Many microalgal industries that started targeting biofuel in the last decade had now adopted parallel business plans focusing on algae by-products application as cosmetic supplements, nutraceuticals, oils, natural color, and animal feed. This review provides the current status and proposes a framework for key supply demand, challenges for cost-effective and sustainable use of water and nutrient. Emphasis is placed on the future industrial market status of value added by products of microalgal biomass. The cost factor for biorefinery process development needs to be addressed before its potential to be exploited for various value-added products with algal biofuel.

Growth of Basil (Ocimum basilicum) in Aeroponics, DRF, and Raft Systems with Effluents of African Catfish (Clarias gariepinus) in Decoupled Aquaponics (s.s.)

Basil (Ocimum basilicum) was cultivated in three hydroponic subsystems (i) a modified commercial aeroponics, (ii) a dynamic root floating (DRF) system, and (iii) a floating raft system in a decoupled aquaponic system in Northern Germany, Mecklenburg–Western Pomerania. For plant nutrition, aquaculture process water from intensive rearing of African catfish (Clarias gariepinus) was used without fertilizer. After 39 days, 16 plant growth parameters were compared, with aeroponics performing significantly better in 11 parameters compared with the DRF, and better compared with the raft in 13 parameters. The economically important leaf wet and dry weight was over 40% higher in aeroponics (28.53 ± 8.74 g; 4.26 ± 1.23 g), but similar in the DRF (20.19 ± 6.57 g; 2.83 ± 0.90 g) and raft (20.35 ± 7.14 g; 2.84 ± 1.04 g). The roots in the DRF grew shorter and thicker; however, this resulted in a higher root dry weight in aeroponics (1.08 ± 0.38 g) compared with the DRF (0.82 ± 0.36 g) and raft (0.67 ± 0.27 g). With optimal fertilizer and system improvement, aquaponic aeroponics (s.s.) could become a productive and sustainable large-scale food production system in the future. Due to its simple construction, the raft is ideal for domestic or semi-commercial use and can be used in areas where water is neither scarce nor expensive. The DRF system is particularly suitable for basil cultivation under hot tropical conditions.

Bioprocess Optimisation for High Cell Density Endoinulinase Production from Recombinant Aspergillus niger.

Endoinulinase gene was expressed in recombinant Aspergillus niger for selective and high-level expression using an exponential fed-batch fermentation. The effects of the growth rate (μ), glucose feed concentration, nitrogen concentration and fungal morphology on enzyme production were evaluated. A recombinant endoinulinase with a molecular weight of 66 kDa was secreted. Endoinulinase production was growth associated at μ> 0.04 h-1, which is characteristic of the constitutive gpd promoter used for the enzyme production. The highest volumetric activity (670 U/ml) was achieved at a growth rate of 93% of μmax (0.07 h-1), while enzyme activity (506 U/ml) and biomass substrate yield (0.043 gbiomassDW/gglucose) significantly decreased at low μ (0.04 h-1). Increasing the feed concentration resulted in high biomass concentrations and viscosity, which necessitated high agitation to enhance the mixing efficiency and oxygen. However, the high agitation and low DO levels (ca. 8% of saturation) led to pellet disruption and growth in dispersed morphology. Enzyme production profiles, product (Yp/s) and biomass (Yx/s) yield coefficients were not affected by feed concentration and morphological change. The gradual increase in the concentration of nitrogen sources showed that, a nitrogen limited culture was not suitable for endoinulinase production in recombinant A. niger. Moreover, the increase in enzyme volumetric activity was still directly related to an increase in biomass concentration. An increase in nitrogen concentration, from 3.8 to 12 g/L, resulted in volumetric activity increase from 393 to 670 U/ml, but the Yp/s (10053 U/gglucose) and Yx/s (0.049 gbiomasDWs/gglucose) did not significantly change. The data demonstrated the potential of recombinant A. niger and high cell density fermentation for the development of large-scale endoinulinase production system.

Two Sides of the Same Coin: A Combination of Archaeometallurgy and Environmental Archaeology to Re-Examine the Hypothesis of Yunnan as the Source of Highly Radiogenic Lead in Early Dynastic China

Bronze Age Shang China is characterized by its large-scale production system and distinctive ritual world. Both are vividly materialized by a large number of bronze ritual vessels with added lead. Whilst a remarkable amount of research effort has been channeled into the trace elemental and lead isotopic analysis of these ritual vessels, and successfully revealed some important fingerprints such as highly radiogenic lead (HRL), there is as yet no consensus on the metal source(s) which supplied the entire bronze production during the Shang period. In addition to the traditional method to look for matching and mismatching between ores and objects, we propose that environmental archaeological studies can provide crucial clues to address some long-standing questions in archaeometallurgy. In the first part of the paper, we attempt to illustrate the potential and complexity of combining these two subjects together. The second part of the paper offers a case study by reviewing the debate on Yunnan as the source of HRL. Synthesis of various lines of evidence published by most recent studies on environmental archaeology, archaeometallurgy, field reports and radiocarbon dating suggests that this hypothesis appears much less likely than previously suspected.

Cell free protein synthesis versus yeast expression – A comparison using insulin as a model protein

Expression of recombinant proteins traditionally require a cellular system to transcribe and translate foreign DNA to a desired protein. The process requires special knowledge of the specific cellular metabolism in use and is often time consuming and labour intensive. A cell free expression system provides an opportunity to express recombinant proteins without consideration of the living cell. Instead, a cell free system relies on either a cellular lysate or recombinant proteins to carry out protein synthesis, increasing overall production speed and ease of handling. The one-pot cell free setup is commonly known as an in vitro transcription/translation reaction (IVTT).Here we focused on a PURE (Protein synthesis Using Recombinant Elements) IVTT system based on recombinant proteins from Escherichia coli. We evaluated the cell free system's ability to express functional insulin analogues compared to Saccharomyces cerevisiae, a well-established system for large scale production of recombinant human insulin and insulin analogues.Significantly, it was found that correct insulin expression and folding was governed by the inherent properties of the primary amino acids sequence of insulin, whereas the eukaryotic features of the expression system apparently play a minor role. The IVTT system successfully produced insulin analogues identical in structure and with similar insulin receptor affinity to those produced by yeast. In conclusion we demonstrate that the PURE IVTT system is highly suited for expressing soluble molecules with higher order features and multiple disulphide bridges.

Optimization of culture conditions for human bone marrow-derived mesenchymal stromal cell expansion in macrocarrier-based Tide Motion system.

With high cell doses required for mesenchymal stromal cell (MSC) clinical trials, there is a need to upgrade technologies that facilitate efficient scale up of MSCs for cell therapy. Conventional expansion with 2D culture vessels becomes the bottleneck when large cell dosages are required. Tide Motion bioreactors offer a robust, scalable platform using BioNOC II macrocarriers developed for the production of adherent cells. We evaluated the growth and expansion of bone marrow-derived MSCs (BM-MSCs) on the macrocarrier-based culture system by optimizing key parameters such as cell seeding densities, culturing conditions, and harvesting procedures to achieve optimal cell growth. BM-MSCs expanded in conventional 2D adherent cultures were seeded into BioNOC II macrocarriers and grown in serum-containing or serum-free medium. BM-MSCs attained a maximum cell density of 0.49 ± 0.07×106 cells/carrier after 12 days of culture in BioNOC II macrocarriers with cell viability > 86% while retaining MSC specific characteristics such as surface marker expression, tri-lineage differentiation potential, immunosuppressive properties, and potency. These results reveal the feasibility of BM-MSC expansion in the scalable macrocarrier-based Tide Motion system both under serum and serum-free conditions and represent an important step for the large-scale production system of BM-MSC based cellular therapies.

Cultivating Microalgae in Desert Conditions: Evaluation of the Effect of Light-Temperature Summer Conditions on the Growth and Metabolism of Nannochloropsis QU130

Temperature and light are two of the most crucial factors for microalgae production. Variations in these factors alter their growth kinetics, macromolecular composition and physiological properties, including cell membrane permeability and fluidity. The variations define the adaptation mechanisms adopted by the microalgae to withstand changes in these environmental factors. In the Qatar desert the temperature varies widely, typically between 10° and 45 °C There are also wide variations in light intensity, with values of over 1500 μmolhν.m−2s−1 in summer. A study of the effects of these thermal and light fluctuations is therefore essential for large-scale outdoor production systems, especially during the summer when temperature and light fluctuations are at their highest. The aim of this work is to study the impact of temperature and light intensity variations as encountered in summer period on the Nannochloropsis QU130 strain, which was selected for its suitability for outdoor cultivation in the harsh conditions of the Qatar desert. It was carried out using lab-scale photobioreactors enabling simulation of both constant and dynamic temperature and light regimes. Biomass productivity, cell morphology and biochemical compositions were examined first in constant conditions, then in typical outdoor cultivation conditions to elucidate the adjustments in cell function in respect of fluctuations. The dynamic light and temperature were shown to have interactive effects. The application of temperature cycles under constant light led to a 13.6% increase in biomass productivity, while a 45% decrease was observed under light and temperature regimes due to the combined stress. In all cases, the results proved that N. sp. QU130 has a high level of adaptation to the wide fluctuations in light and temperature stress. This was shown through its ability to easily change its physiology (cell size) and metabolic process in response to different cultivation conditions.