3D bioprinting – Flow cytometry as analytical strategy for 3D cell structures

Gretzinger, Sarah; Beckert, Nicole; Gleadall, Andrew; Lee-Thedieck, Cornelia; Hubbuch, Jürgen

doi:10.1016/j.bprint.2018.e00023

Cited by 9 publications

(12 citation statements)

References 47 publications

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“…Firstly, they provide different levels of detail for the mesh. Liravi et al [40] used a very specific mesh generation with a remeshing procedure developed by Wilkes et al [42] to properly generate droplets. On the contrary, Samanipour et al [41] did not even mention the mesh they generated for their simulations.…”

Section: Introductionmentioning

confidence: 99%

Bioink Temperature Influence on Shear Stress, Pressure and Velocity Using Computational Simulation

et al. 2020

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Bioinks are usually cell-laden hydrogels widely studied in bioprinting performing experimental tests to tune their rheological properties, thus increasing research time and development costs. Computational Fluids Dynamics (CFD) is a powerful tool that can minimize iterations and costs simulating the material behavior using parametric changes in rheological properties under testing. Additionally, most bioinks have specific functionalities and their properties might widely change with temperature. Therefore, commercial bioinks are an excellent way to standardize bioprinting process, but they are not analyzed in detail. Therefore, the objective of this work is to study how three temperatures of the Cellink Bioink influence shear stress pressure and velocity through computational simulation. A comparison of three conical nozzles (20, 22, and 25G) for each temperature has been performed. The results show that shear stress, pressure, and velocity vary in negligible ranges for all combinations. Although these ranges are small and define a good thermo-responsive bioink, they do not generate a filament on the air and make drops during extrusion. In conclusion, this bioink provides a very stable behavior with low shear stress, but other bioprinting parameters must be set up to get a stable filament width.

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Section: Introductionmentioning

confidence: 99%

Bioink Temperature Influence on Shear Stress, Pressure and Velocity Using Computational Simulation

et al. 2020

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“…The reproducibility as well as the significance of the results regarding cell viability can be increased by the implementation of automated image processing workflows, as well [25,26]. A further option to quantify several thousand cells is flow cytometry as already tested in a few studies in the field [27,28]. The lack of standardized printing methodologies as well as techniques for the evaluation of the printing performance of bioinks presents a limitation in the development of formulations as a comparison within and between laboratories.…”

Section: Introductionmentioning

confidence: 99%

Analytics in Extrusion-Based Bioprinting: Standardized Methods Improving Quantification and Comparability of the Performance of Bioinks

2023

Self Cite

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Three-dimensional bioprinting and especially extrusion-based printing as a most frequently employed method in this field is constantly evolving as a discipline in regenerative medicine and tissue engineering. However, the lack of relevant standardized analytics does not yet allow an easy comparison and transfer of knowledge between laboratories regarding newly developed bioinks and printing processes. This work revolves around the establishment of a standardized method, which enables the comparability of printed structures by controlling for the extrusion rate based on the specific flow behavior of each bioink. Furthermore, printing performance was evaluated by image-processing tools to verify the printing accuracy for lines, circles, and angles. In addition, and complementary to the accuracy metrics, a dead/live staining of embedded cells was performed to investigate the effect of the process on cell viability. Two bioinks, based on alginate and gelatin methacryloyl, which differed in 1% (w/v) alginate content, were tested for printing performance. The automated image processing tool reduced the analytical time while increasing reproducibility and objectivity during the identification of printed objects. During evaluation of the processing effect of the mixing of cell viability, NIH 3T3 fibroblasts were stained and analyzed after the mixing procedure and after the extrusion process using a flow cytometer, which evaluated a high number of cells. It could be observed that the small increase in alginate content made little difference in the printing accuracy but had a considerable strong effect on cell viability after both processing steps.

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“…The natural distribution of cells throughout the 3D environment, as well as changes within the population of encapsulated 3D cell models, demand a thorough data-collecting strategy to obtain the necessary information. Furthermore, in 3D cell models embedded within a hydrogel environment, the hydrogel itself may interfere with or affect assay readouts or imaging techniques [31,34]. Therefore, standardising, and validating 3D cell culture model assay approaches to provide reproducibility, automation, and high throughput analysis is much needed for further advancements in 3D cell culture models [28].…”

Section: Introductionmentioning

confidence: 99%

Flow cytometry as an analytical method of drug-induced apoptosis in 3D bioprinted melanoma cells

Villiers

Plessis

2023

Biomed. Mater.

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Three-dimensional (3D) cell culture systems have gained increasing interest in drug discovery and tissue engineering due to its inherent advantages in providing more physiologically relevant information and more predictive data for in vivo tests. Along with the development of more physiologically relevant 3D cell culture models, researchers bear the responsibility to validate new cell assay techniques capable of measuring and evaluating constructs that are physically larger and more complex compared to two- dimensional (2D) cell cultures. It is important to note that assays based on monolayer cultures may be insufficient for the use in 3D cell cultures models. In this study we firstly fabricated a 3D bioprinted hydrogel melanoma scaffold. This was used to validate a flow cytometry-based analytical method as a tool for 3D bioprinted structures to assess drug-induced apoptosis. The results indicated high robustness, reproducibility and sensitivity of the flow cytometric method established on the 3D cell-laden A375 melanoma hydrogel scaffolds. Over and above this, it was possible to determine the effect of etoposide on A375 melanoma cells using Annexin V and PI apoptosis assay.

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3D bioprinting – Flow cytometry as analytical strategy for 3D cell structures

Cited by 9 publications

References 47 publications

Bioink Temperature Influence on Shear Stress, Pressure and Velocity Using Computational Simulation

Bioink Temperature Influence on Shear Stress, Pressure and Velocity Using Computational Simulation

Analytics in Extrusion-Based Bioprinting: Standardized Methods Improving Quantification and Comparability of the Performance of Bioinks

Flow cytometry as an analytical method of drug-induced apoptosis in 3D bioprinted melanoma cells

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