Quantitative Validation of the Presto Blue™ Metabolic Assay for Online Monitoring of Cell Proliferation in a 3D Perfusion Bioreactor System

Sonnaert, Maarten; Papantoniou, Ioannis; Luyten, Frank P.; Schrooten, Jan

doi:10.1089/ten.tec.2014.0255

Cited by 52 publications

(42 citation statements)

References 47 publications

(95 reference statements)

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“…The detection of cell viability depends on the functions of the mitochondrial enzymes within the live cells which reacted with the rezasurin compound [37]- [39] The fluorescent reduced formed of rezasurin changed the color of the cell media and could be measured by both fluorescence and absorbance [40]. A quantitative study stated that Prestoblue gives a stronger fluorescent signal than Alamar blue assay [41]. Prestoblue assay is a ready-to-use assay as it only needed one single step compared to MTT and LDH release assays [39].…”

Section: Resultsmentioning

confidence: 99%

Evaluation of Bismuth Oxide Nanoparticles as Radiosensitizer for Megavoltage Radiotherapy

Sisin¹,

Abidin²,

Amir³

et al. 2019

Int. J. Adv. Sci. Eng. Inf. Technol.

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Metal-based nanoparticles such as gold, silver, platinum, and bismuth have been widely investigated for radiotherapeutic application. Basic understanding of the cellular interaction of the nanoparticles with the biological materials is crucial to ensure future clinical use. In this study, the cytotoxicity, cellular uptake, and generation of reactive oxygen species (ROS) induced by BiONPs were investigated prior elucidating the feasibility of BiONPs for radiotherapy application using megavoltage photon and electron beams. The BiONPs of diameter sizes 60, 70, 80 and 90 nm at concentrations within a range of 0.5 to 0.00005 mMol/L were tested on MCF-7, MDA-MB-231, and NIH/3T3 cells lines. The cytotoxicity results exhibit minimal cell death constituting less than 20 % of mortality on average. The ROS generation by BiONPs alone is found to be negligible as the ROS levels were slightly lower and higher than 100% of positive control. The increment of cellular nanoparticles uptake from a range of 1.50 % to 34.10 % indicates that BiONPs were internalized and bound to the surface of the cells. Sequencing from the results, 60 nm BiONPs are found to be the most suitable to be applied as a radiosensitizer in radiotherapy. Sensitization enhancement ratio (SER) quantified on MCF-7 cells demonstrated the highest enhancement from the highest concentration of BiONPs with SER of 2.29 and 1.42, for 10 MV photon beam and 6 MeV electron beam, respectively. In contrast to ROS production without radiation, the ROS induced from radiotherapy beams were found to be dose-dependent and play significant roles in radiosensitization effect. In conclusion, BiONPs could improve clinical radiotherapy, and further radiobiological characterization is crucial for future clinical translation.

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Section: Resultsmentioning

confidence: 99%

Evaluation of Bismuth Oxide Nanoparticles as Radiosensitizer for Megavoltage Radiotherapy

Sisin¹,

Abidin²,

Amir³

et al. 2019

Int. J. Adv. Sci. Eng. Inf. Technol.

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“…For 3D perfusion bioreactor cell expansion, additive manufactured Ti6Al4V scaffolds (Ø = 6 mm, h = 6 mm) [ 34 , 35 ] were used as described before [ 11 , 12 , 32 , 36 ]. The total volume of the scaffolds was 166 ± 3 mm 3 , the available volume 130 ± 5 mm 3 and the available surface 7.5 ± 0.6 mm 2 as determined with μCT [ 37 ].…”

Section: Methodsmentioning

confidence: 99%

“…The total volume of the scaffolds was 166 ± 3 mm 3 , the available volume 130 ± 5 mm 3 and the available surface 7.5 ± 0.6 mm 2 as determined with μCT [ 37 ]. Scaffolds were cleaned and prepared for experiments as described before [ 11 , 12 , 32 , 36 ].…”

Section: Methodsmentioning

confidence: 99%

“…Prior to the 3D culture experiments cells were harvested at passage 6 using Triple Express (Life Technologies) and drop-seeded onto the scaffolds at a final density of 120,000 cells per scaffold [ 11 , 12 , 32 , 36 ]. Scaffold-cell constructs were incubated overnight in standard culture conditions resulting in an initial cell density of 17,100 cells/cm 2 or 700,000 cells/cm 3 .…”

Section: Methodsmentioning

confidence: 99%

“…In this work we expanded human periosteum derived cells (hPDCs) in a 3D flow-through perfusion bioreactor [ 11 ] and monitored their growth non-destructively using the Presto Blue metabolic assay [ 32 ]. Upon confluence, as was confirmed by contrast enhanced X-ray nano-computed tomography (CE-nanoCT), a range of cell recovery reagents was assessed.…”

Section: Introductionmentioning

confidence: 99%

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Bioreactor-Based Online Recovery of Human Progenitor Cells with Uncompromised Regenerative Potential: A Bone Tissue Engineering Perspective

et al. 2015

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The use of a 3D perfusion culture environment for stem cell expansion has been shown to be beneficial for maintenance of the original cell functionality but due to several system inherent characteristics such as the presence of extracellular matrix, the continued development and implementation of 3D perfusion bioreactor technologies is hampered. Therefore, this study developed a methodology for harvesting a progenitor cell population from a 3D open porous culture surface after expansion in a perfusion bioreactor and performed a functional characterization of the expanded cells. An initial screening showed collagenase to be the most interesting reagent to release the cells from the 3D culture surface as it resulted in high yields without compromising cell viability. Subsequently a Design of Experiment approach was used to obtain optimized 3D harvest conditions by assessing the interplay of flow rate, collagenase concentration and incubation time on the harvest efficiency, viability and single cell fraction. Cells that were recovered with the optimized harvest protocol, by perfusing a 880 U/ml collagenase solution for 7 hours at a flow rate of 4 ml/min, were thereafter functionally analyzed for their characteristics as expanded progenitor cell population. As both the in vitro tri-lineage differentiation capacity and the in vivo bone forming potential were maintained after 3D perfusion bioreactor expansion we concluded that the developed seeding, culture and harvest processes did not significantly compromise the viability and potency of the cells and can contribute to the future development of integrated bioprocesses for stem cell expansion.

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Tissue‐Adaptive Materials with Independently Regulated Modulus and Transition Temperature

et al. 2020

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Various types of clinical complications may emerge when a medical device (catheter, microelectrode, or microneedle patch) is deployed to the human body. It has been widely recognized that mechanical mismatch between an implant and a host tissue is one of the leading factors for adverse effects such as irritation and inflammation, which is evident in orthopedic, [1] neural, [2,3] and reconstructive implants. [4] It is generally desired that an implant material precisely matches Young's modulus of the surrounding tissue that may range between E ≈ 10 2 Pa of supersoft brain and adipose tissues to E ≈ 10 5 Pa of muscle and skin. However, forceful implantation of biomedical devices such as piercing, insertion or twisting impose significant mechanical stresses on the device. [5-8] Therefore a certain rigidity is desired to prevent device failure, minimize tissue damage, and simplify storage and handling procedures. [9,10] This presents an oxymoronic challenge for an ideal implant: designing an adaptive material that readily penetrates tissue, but transitions into a tissue-soft implant upon insertion into the body (Figure 1A). Thermoplastics are one of the most commonly used materials possessing a well-defined transition between hard and soft states with Young's moduli of E hard ≈ 1 GPa and E soft ≈ 1 MPa, respectively (Figure 1B, Table S1, Supporting Information). However, conventional thermoplastic materials have two problems: i) E soft ≈ 1 MPa is significantly higher than soft tissue modulus ranging within 10 2-10 5 Pa and ii) softening (melting or glass) transition temperature of high molecular weight polymers cannot be tuned without altering their chemical composition. It is even more challenging to adjust the mechanical and thermal properties independently of each other. The softness issue alone could be resolved by using different types of bioinspired polymeric gels. [14-17] However, not only it is difficult to prepare gels with a sharp GPa-to-kPa transition (Figure 1C, Table S2, Supporting Information), their solvent fraction instigates uncontrolled leaching inside the body, [13] which causes various health risks. The second problem can be addressed via copolymerization of monomers with different melting temperatures. [18-20] However, this involves chemical composition and biocompatibility variation and yet, copolymer modulus is limited by chain entanglements to >10 5 Pa. [21,22] The ability of living species to transition between rigid and flexible shapes represents one of their survival mechanisms, which has been adopted by various human technologies. Such transition is especially desired in medical devices as rigidity facilitates the implantation process, while flexibility and softness favor biocompatibility with surrounding tissue. Traditional thermoplastics cannot match soft tissue mechanics, while gels leach into the body and alter their properties over time. Here, a single-component system with an unprecedented drop of Young's modulus by up to six orders of magnitude from the GPa to kPa level at a control...

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Quantitative Validation of the Presto Blue™ Metabolic Assay for Online Monitoring of Cell Proliferation in a 3D Perfusion Bioreactor System

Cited by 52 publications

References 47 publications

Evaluation of Bismuth Oxide Nanoparticles as Radiosensitizer for Megavoltage Radiotherapy

Evaluation of Bismuth Oxide Nanoparticles as Radiosensitizer for Megavoltage Radiotherapy

Bioreactor-Based Online Recovery of Human Progenitor Cells with Uncompromised Regenerative Potential: A Bone Tissue Engineering Perspective

Tissue‐Adaptive Materials with Independently Regulated Modulus and Transition Temperature

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