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Please use this identifier to cite or link to this item: https://doi.org/10.21256/zhaw-25998
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dc.contributor.authorBono, Epifania-
dc.contributor.authorSheard, Jonathan-
dc.contributor.authorMikkonen, Piia-
dc.contributor.authorRimann, Markus-
dc.date.accessioned2022-11-11T09:57:01Z-
dc.date.available2022-11-11T09:57:01Z-
dc.date.issued2022-09-
dc.identifier.urihttps://digitalcollection.zhaw.ch/handle/11475/25998-
dc.description.abstractIntroduction: It is recognized that screening of drugs on 2D models is unable to precisely select clinically active medicaments; therefore, 3D culture systems are emerging and show potential for better simulating the in vivo tumour microenvironment and for eliminating the species differences to allow drug testing directly in human systems before drugs move into clinical trials. The purpose of this study was to automate the production and cultivation of human primary osteogenic sarcoma cells line, SaOS-2, in scaffold-based, multiple spheroids in GrowDex® (GDS) and scaffold-free systems, single spheroids (SS) for high-throughput screening. Methods: For scaffold-based models, SAOS-2 were embedded in nanofibrillar cellulose hydrogel, GrowDex®, in flat ultra-low attachment (ULA) 96-well plates while for the scaffold-free system cells were let to form spheroids in U-bottom ULA 96-well plates. Experiments were conducted on the Fluent® 780 automation workstation. The scripts for automated model production and media exchange were established in the FluentControlTM. Technical parameters such as aspiration and dispensing speed as well as XYZ dispensing positions were empirically defined and optimized i) to allow production of stable models, ii) to avoid disturbing during the medium exchange neither the GDS nor the GrowDex structure/shape, iii) to avoid SS displacement and aspiration. GDS and SS models were cultivated up to 11 days and were characterized for their viability (ATP assay), morphology and size (MTT assay). Dose-response tests with taxol and doxorubicin were carried out for both system types. Drugs were automated dispensed on the 3D models at day 4 for a treatment time of 72h. Cells were analysed for cell viability, measuring ATP levels, morphology, and size. Results: The established scripts allowed the formation of multiple spheroids in GrowDex as well as the aggregation of SAOS-2 in single spheroids in the U-bottom ULA plates. 70% of medium could be successfully automated exchanged maintaining unaltered the shape and the position of the models in the wells. GDS and SS remained stable for up to 11 days and increased in size over time, showing a similar growth rate. Viable GDS populated the entire model at different Z values with a compact morphology, parameter that characterized the SS too. GDS showed, as expected, a wide size distribution while SS were bigger and more homogenous in size in comparison to GDS (SS area 10-fold bigger than GDS area). SAOS-2 responded in both systems to taxol and doxorubicin, showing higher IC50 values for GDS compared to SS. Taxol (fig.1) and doxorubicin were 3.5- and 4.5-fold more potent in SS than in GDS. Collapsed morphology with a viable and compact core and a loose cell layer around border at high drugs concentrations characterized exclusively SS. Discussion & Conclusions: The automation protocols were successfully established allowing the reproducible production and maintenance of GrowDex multiple spheroids and scaffold-free models. Although its viscosity, GrowDex is automation compatible, and the results obtained in this project shows its high potential for high-throughput drug screening. Acknowledgements: The ZHAW authors would like to thank the Tecan for the technical support and the consumables, and the UPM Biomedicals for its contribution.de_CH
dc.language.isoende_CH
dc.publisherZHAW Zürcher Hochschule für Angewandte Wissenschaftende_CH
dc.rightsLicence according to publishing contractde_CH
dc.subjectAutomationde_CH
dc.subjectCellulose-based scaffoldde_CH
dc.subjectDrug screeningde_CH
dc.subjectSpheroidde_CH
dc.subject.ddc610.28: Biomedizin, Biomedizinische Technikde_CH
dc.titleAutomation of 3D cell culture using cellulose-based scaffoldsde_CH
dc.typeKonferenz: Posterde_CH
dcterms.typeTextde_CH
zhaw.departementLife Sciences und Facility Managementde_CH
zhaw.organisationalunitInstitut für Chemie und Biotechnologie (ICBT)de_CH
zhaw.publisher.placeWinterthurde_CH
dc.identifier.doi10.21256/zhaw-25998-
zhaw.conference.detailsBiointerfaces International Conference (BIC), Zurich, Switzerland, 12-15 September 2022de_CH
zhaw.funding.euNode_CH
zhaw.originated.zhawYesde_CH
zhaw.publication.statuspublishedVersionde_CH
zhaw.publication.reviewNot specifiedde_CH
zhaw.webfeed3D Gewebe und Biofabrikationde_CH
zhaw.author.additionalNode_CH
zhaw.display.portraitYesde_CH
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Bono, E., Sheard, J., Mikkonen, P., & Rimann, M. (2022, September). Automation of 3D cell culture using cellulose-based scaffolds. Biointerfaces International Conference (BIC), Zurich, Switzerland, 12-15 September 2022. https://doi.org/10.21256/zhaw-25998
Bono, E. et al. (2022) ‘Automation of 3D cell culture using cellulose-based scaffolds’, in Biointerfaces International Conference (BIC), Zurich, Switzerland, 12-15 September 2022. Winterthur: ZHAW Zürcher Hochschule für Angewandte Wissenschaften. Available at: https://doi.org/10.21256/zhaw-25998.
E. Bono, J. Sheard, P. Mikkonen, and M. Rimann, “Automation of 3D cell culture using cellulose-based scaffolds,” in Biointerfaces International Conference (BIC), Zurich, Switzerland, 12-15 September 2022, Sep. 2022. doi: 10.21256/zhaw-25998.
BONO, Epifania, Jonathan SHEARD, Piia MIKKONEN und Markus RIMANN, 2022. Automation of 3D cell culture using cellulose-based scaffolds. In: Biointerfaces International Conference (BIC), Zurich, Switzerland, 12-15 September 2022. Conference poster. Winterthur: ZHAW Zürcher Hochschule für Angewandte Wissenschaften. September 2022
Bono, Epifania, Jonathan Sheard, Piia Mikkonen, and Markus Rimann. 2022. “Automation of 3D Cell Culture Using Cellulose-Based Scaffolds.” Conference poster. In Biointerfaces International Conference (BIC), Zurich, Switzerland, 12-15 September 2022. Winterthur: ZHAW Zürcher Hochschule für Angewandte Wissenschaften. https://doi.org/10.21256/zhaw-25998.
Bono, Epifania, et al. “Automation of 3D Cell Culture Using Cellulose-Based Scaffolds.” Biointerfaces International Conference (BIC), Zurich, Switzerland, 12-15 September 2022, ZHAW Zürcher Hochschule für Angewandte Wissenschaften, 2022, https://doi.org/10.21256/zhaw-25998.


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