Microarrays for the scalable production of metabolically relevant tumour spheroids: a tool for modulating chemosensitivity traits
| Autor: | Heike Hardelauf, Joanna Stewart, Jan G. Hengstler, Wiebke Schormann, Joachim Franzke, Ya-Yu Chiang, Jean-Philippe Frimat, Jonathan West, Cristina Cadenas, Peter Lampen, Leoni A. Kunz-Schughart |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Cell cycle checkpoint
Biomedical Engineering Bioengineering 02 engineering and technology Biology Biochemistry 03 medical and health sciences Colon carcinoma ddc:570 acid-phosphatase assay culture-system hepatocyte spheroids topoisomerase-i cell viability growth microfabrication differentiation camptothecin generation Spheroids Cellular Tumour spheroid ddc:012 Humans Viability assay Dimethylpolysiloxanes 030304 developmental biology 0303 health sciences Drug discovery Tumor biology Cell Cycle Spheroid General Chemistry 021001 nanoscience & nanotechnology Microarray Analysis Nylons Saure Phosphatase Untersuchung Zellkultursystem Hepatozyt Sphäroide Topoisomerase Zellviabilität Wachstum Mikrofabrikation Differenzierung Camptothecin embryonic structures Colonic Neoplasms Biophysics Regression Analysis DNA microarray 0210 nano-technology HT29 Cells Biomedical engineering |
| Zdroj: | Lab Chip; Vol 11 Lab Chip LAB on a chip 2011;11:419–428, ISSN: 1473-0197 |
| ISSN: | 1473-0197 |
| DOI: | 10.1039/C0LC00089B |
| Popis: | We report the use of thin film poly(dimethylsiloxane) (PDMS) prints for the arrayed mass production of highly uniform 3-D human HT29 colon carcinoma spheroids. The spheroids have an organotypic density and, as determined by 3-axis imaging, were genuinely spherical. Critically, the array density impacts growth kinetics and can be tuned to produce spheroids ranging in diameter from 200 to 550 µm. The diffusive limit of competition for media occurred with a pitch of ≥1250 µm and was used for the optimal array-based culture of large, viable spheroids. During sustained culture mass transfer gradients surrounding and within the spheroids are established, and lead to growth cessation, altered expression patterns and the formation of a central secondary necrosis. These features reflect the microenvironment of avascularised tumours, making the array format well suited for the production of model tumours with defined sizes and thus defined spatio-temporal pathophysiological gradients. Experimental windows, before and after the onset of hypoxia, were identified and used with an enzyme activity-based viability assay to measure the chemosensitivity towards irinotecan. Compared to monolayer cultures, a marked reduction in the drug efficacy towards the different spheroid culture states was observed and attributed to cell cycle arrest, the 3-D character, scale and/or hypoxia factors. In summary, spheroid culture using the array format has great potential to support drug discovery and development, as well as tumour biology research. Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich. |
| Databáze: | OpenAIRE |
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