bioreactor system
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2022 ◽  
Vol 9 (1) ◽  
pp. 39
Jihyoung Choi ◽  
Sanjana Mathew ◽  
Sabrina Oerter ◽  
Antje Appelt-Menzel ◽  
Jan Hansmann ◽  

Electrochemical impedance spectroscopy (EIS) is a noninvasive, reliable, and efficient method to analyze the barrier integrity of in vitro tissue models. This well-established tool is used most widely to quantify the transendothelial/epithelial resistance (TEER) of Transwell-based models cultured under static conditions. However, dynamic culture in bioreactors can achieve advanced cell culture conditions that mimic a more tissue-specific environment and stimulation. This requires the development of culture systems that also allow for the assessment of barrier integrity under dynamic conditions. Here, we present a bioreactor system that is capable of the automated, continuous, and non-invasive online monitoring of cellular barrier integrity during dynamic culture. Polydimethylsiloxane (PDMS) casting and 3D printing were used for the fabrication of the bioreactors. Additionally, attachable electrodes based on titanium nitride (TiN)-coated steel tubes were developed to perform EIS measurements. In order to test the monitored bioreactor system, blood–brain barrier (BBB) in vitro models derived from human-induced pluripotent stem cells (hiPSC) were cultured for up to 7 days. We applied equivalent electrical circuit fitting to quantify the electrical parameters of the cell layer and observed that TEER gradually decreased over time from 2513 Ω·cm2 to 285 Ω·cm2, as also specified in the static control culture. Our versatile system offers the possibility to be used for various dynamic tissue cultures that require a non-invasive monitoring system for barrier integrity.

Paul Ritter ◽  
Aijia Cai ◽  
Barbara Reischl ◽  
Maren Fiedler ◽  
Gerhard Prol ◽  

2021 ◽  
Mariusz Pożoga ◽  
Dawid Olewnicki ◽  
Elżbieta Wójcik-Gront

Abstract The aim of this study was to propose an efficient method of Pennisetum x advena ‘Rubrum’ micropropagation. Agar cultures with MS medium supplemented with BAP in various concentrations (0.5 mg/L-2 mg/L) and a temporary immersion bioreactor system (TIS) with liquid medium MS with an addition of 1 mg/L BAP were used. For rooting ½ MS medium with different auxin combinations (IBA, NAA) and activated charcoal was utilized. The most efficient method turned out to be TIS which produced 36.9 new plants in four weeks. The seedlings were slender in shape, bright green in colour with no signs of hyperhydricity. The most suitable agar medium produced 19.5 new plants in an eight week period. Rooting should be carried on ½ MS supplemented with 0.5 mg/L IBA and 0.5 mg/L NAA with an 84% rooting rate. The addition of activated charcoal inhibited rooting.

2021 ◽  
Vol 12 ◽  
Brendon Herring ◽  
Samuel Jang ◽  
Jason Whitt ◽  
Kayla Goliwas ◽  
Zviadi Aburjania ◽  

Few models exist for studying neuroendocrine tumors (NETs), and there are mounting concerns that the currently available array of cell lines is not representative of NET biology. The lack of stable patient-derived NET xenograft models further limits the scientific community’s ability to make conclusions about NETs and their response to therapy in patients. To address these limitations, we propose the use of an ex vivo 3D flow-perfusion bioreactor system for culturing and studying patient-derived NET surrogates. Herein, we demonstrate the utility of the bioreactor system for culturing NET surrogates and provide methods for evaluating the efficacy of therapeutic agents on human NET cell line xenograft constructs and patient-derived NET surrogates. We also demonstrate that patient-derived NET tissues can be propagated using the bioreactor system and investigate the near-infrared (NIR) dye IR-783 for its use in monitoring their status within the bioreactor. The results indicate that the bioreactor system and similar 3D culture models may be valuable tools for culturing patient-derived NETs and monitoring their response to therapy ex vivo.

Alexander P. J. Ebbing ◽  
Gregory S. Fivash ◽  
Ronald Pierik ◽  
Tjeerd J. Bouma ◽  
Jacco C. Kromkamp ◽  

AbstractThe future of large-scale kelp aquaculture is standing at a crossroad, with the diverging paths being characterized by two fundamentally different cultivation methods that differ on how well gametophyte reproduction can be controlled. The cultivation method that does not directly control gametophyte reproduction is more widely utilized at the moment, but interest in better controlling gametophyte reproduction is growing steadily. Here, we validate a bioreactor system that overcomes a number of implementation challenges for this controlled reproductive method, expanding the possibility of clonal gametophyte cultivation outside of expensive laboratory settings. The main goals of this system include (i) the maintenance of clean gametophyte clonal cultures in non-sterile environments over prolonged periods of time, (ii) the production of large numbers of juvenile sporophytes, and (iii) effective transportation of gametophytes and sporophytes. The “SeaCoRe system” consists out of three parts that correspond to these three challenges: (1) clone-reactors, (2) a clone-inducer, and (3) a transporter. The validation of the system showed that delayed Saccharina latissima and Alaria esculenta gametophytes can grow reliably for 75 days in the clone-reactors. Initial gametophyte densities of 0.4 mg DW and 0.6 mg DW gametophtyes mL−1 were optimal for S. latissima and A. esculenta, resulting in reproductive successes of 604 and 422 sporophytes mL−1, respectively. Lastly, gametophyte transport was simulated, with high reproductive success still achieved within 19 days in ~ 20 °C environments. The SeaCoRe system helps unlock the full potential of large-scale kelp cultivation using multiannual delayed clonal.

Yifan Yuan ◽  
Katherine L. Leiby ◽  
Allison M. Greaney ◽  
Micha Sam Brickman Raredon ◽  
Hong Qian ◽  

The development of an in vitro system for the study of lung vascular disease is critical to understanding human pathologies. Conventional culture systems fail to fully recapitulate native microenvironmental conditions and are typically limited in their ability to represent human pathophysiology for the study of disease and drug mechanisms. Whole organ decellularization provides a means to developing a construct that recapitulates structural, mechanical, and biological features of a complete vascular structure. Here, we developed a culture protocol to improve endothelial cell coverage in whole lung scaffolds and used single-cell RNA-sequencing analysis to explore the impact of decellularized whole lung scaffolds on endothelial phenotypes and functions in a biomimetic bioreactor system. Intriguingly, we found that the phenotype and functional signals of primary pulmonary microvascular revert back—at least partially—toward native lung endothelium. Additionally, human induced pluripotent stem cell-derived endothelium cultured in decellularized lung systems start to gain various native human endothelial phenotypes. Vascular barrier function was partially restored, while small capillaries remained patent in endothelial cell-repopulated lungs. To evaluate the ability of the engineered endothelium to modulate permeability in response to exogenous stimuli, lipopolysaccharide (LPS) was introduced into repopulated lungs to simulate acute lung injury. After LPS treatment, proinflammatory signals were significantly increased and the vascular barrier was impaired. Taken together, these results demonstrate a novel platform that recapitulates some pulmonary microvascular functions and phenotypes at a whole organ level. This development may help pave the way for using the whole organ engineering approach to model vascular diseases.

2021 ◽  
Vol 11 (1) ◽  
Rupak Dua ◽  
Hugh Jones ◽  
Philip C. Noble

AbstractRecent advances in materials and manufacturing processes have allowed the fabrication of intricate implant surfaces to facilitate bony attachment. However, refinement and evaluation of these new design strategies are hindered by the cost and complications of animal studies, particularly during early iterations in the development process. To address this problem, we have previously constructed and validated an ex-vivo bone bioreactor culture system that can maintain the viability of bone samples for an extended period ex-vivo. In this study, we investigated the mineralization of a titanium wire mesh scaffold under both static and dynamic culturing using our ex vivo bioreactor system. Thirty-six cancellous bone cores were harvested from bovine metatarsals at the time of slaughter and divided into five groups under the following conditions: Group 1) Isolated bone cores placed in static culture, Group 2) Unloaded bone cores placed in static culture in contact with a fiber-mesh metallic scaffold, Group 3) Bone cores placed in contact with a fiber-mesh metallic scaffold under the constant pressure of 150 kPa, Group 4) Bone core placed in contact with a fiber-mesh metallic scaffold and exposed to cyclic loading with continuous perfusion flow of media within the ex-vivo culture system and Group 5) Bone core evaluated on Day 0 to serve as a positive control for comparison with all other groups at weeks 4 and 7. Bone samples within Groups 1–4 were incubated for 4 and 7 weeks and then evaluated using histological examination (H&E) and the Live-Dead assay (Life Technologies). Matrix deposits on the metallic scaffolds were examined with scanning electron microscopy (SEM), while the chemical composition of the matrix was measured using energy-dispersive x-ray spectroscopy (EDX). We found that the viability of bone cores was maintained after seven weeks of loading in our ex vivo system. In addition, SEM images revealed crystallite-like structures on the dynamically loaded metal coupons (Group 4), corresponding to the initial stages of mineralization. EDX results further confirmed the presence of carbon at the interface and calcium phosphates in the matrix. We conclude that a bone bioreactor can be used as an alternate tool for in-vivo bone ingrowth studies of new implant surfaces or coatings.

Energies ◽  
2021 ◽  
Vol 14 (21) ◽  
pp. 6885
Seongwon Im ◽  
Mo-Kwon Lee ◽  
Alsayed Mostafa ◽  
Om Prakash ◽  
Kyeong-Ho Lim ◽  

In a lab-scale bioreactor system, (20 L of effective volume in our study) controlling a constant temperature inside bioreactor with a total volume 25 L is a simple process, whereas it is a complicated process in the actual full-scale system. There might exist a localized temperature difference inside the reactor, affecting bioenergy yield. In the present work, the temperature at the middle layer of bioreactor was controlled at 35 °C, while the temperature at top and bottom of bioreactor was controlled at 35 ± 0.1, ±1.5, ±3.0, and ±5.0 °C. The H2 yield of 1.50 mol H2/mol hexoseadded was achieved at ±0.1 and ±1.5 °C, while it dropped to 1.27 and 0.98 mol H2/mol hexoseadded at ±3.0 and ±5.0 °C, respectively, with an increased lactate production. Then, the reactor with automatic agitation speed control was operated. The agitation speed was 10 rpm (for 22 h) under small temperature difference (<±1.5 °C), while it increased to 100 rpm (for 2 h) when the temperature difference between top and bottom of reactor became larger than ±1.5 °C. Such an operation strategy helped to save 28% of energy requirement for agitation while producing a similar amount of H2. This work contributes to facilitating the upscaling of the dark fermentation process, where appropriate agitation speed can be controlled based on the temperature difference inside the reactor.

Molecules ◽  
2021 ◽  
Vol 26 (20) ◽  
pp. 6270
Lan Thi Tran ◽  
Kuaanan Techato ◽  
Van Bon Nguyen ◽  
San-Lang Wang ◽  
Anh Dzung Nguyen ◽  

The purpose of this study was to reuse cassava wastewater (CW) for scaled-up production, via the fermentation of prodigiosin (PG), and to conduct an evaluation of its bioactivities. PG was produced at the yield of high 6150 mg/L in a 14 L-bioreactor system, when the designed novel medium (7 L), containing CW and supplemented with 0.25% casein, 0.05% MgSO4, and 0.1% K2HPO4, was fermented with Serratia marcescens TNU01 at 28 °C in 8 h. The PG produced and purified in this study was assayed for some medical effects and showed moderate antioxidant, high anti-NO (anti-nitric oxide), and potential α-glucosidase inhibitory activities. Notably, PG was first reported as a novel effective α-glucosidase inhibitor with a low IC50 value of 0.0183 µg/mL. The commercial anti-diabetic drug acarbose was tested for comparison and had a lesser effect with a high IC50 value of 328.4 µg/mL, respectively. In a docking study, the cation form of PG (cation-PG) was found to bind to the enzyme α-glucosidase by interacting with two prominent amino acids, ASP568 and PHE601, at the binding site on the target enzyme, creating six linkages and showing a better binding energy score (−14.6 kcal/mol) than acarbose (−10.5 kcal/mol). The results of this work suggest that cassava wastewater can serve as a low-cost raw material for the effective production of PG, a potential antidiabetic drug candidate.

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