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@article{1708596,
author = {Basu, Amrita and Dydowiczová, Aneta and Trosko, James E. and Bláha, Luděk and Babica, Pavel},
article_number = {8},
doi = {http://dx.doi.org/10.1080/15376516.2020.1800881},
keywords = {Multicellular spheroids; 3D cell cultures; hepatotoxicity; in vitro toxicity testing},
language = {eng},
issn = {1537-6516},
journal = {Toxicology Mechanisms and Methods},
title = {Ready to go 3D? A semi-automated protocol for microwell spheroid arrays to increase scalability and throughput of 3D cell culture testing},
url = {https://www.tandfonline.com/doi/abs/10.1080/15376516.2020.1800881?journalCode=itxm20},
volume = {30},
year = {2020}
}
TY - JOUR
ID - 1708596
AU - Basu, Amrita - Dydowiczová, Aneta - Trosko, James E. - Bláha, Luděk - Babica, Pavel
PY - 2020
TI - Ready to go 3D? A semi-automated protocol for microwell spheroid arrays to increase scalability and throughput of 3D cell culture testing
JF - Toxicology Mechanisms and Methods
VL - 30
IS - 8
SP - 590-604
EP - 590-604
PB - Taylor and Francis Ltd.
SN - 15376516
KW - Multicellular spheroids
KW - 3D cell cultures
KW - hepatotoxicity
KW - in vitro toxicity testing
UR - https://www.tandfonline.com/doi/abs/10.1080/15376516.2020.1800881?journalCode=itxm20
L2 - https://www.tandfonline.com/doi/abs/10.1080/15376516.2020.1800881?journalCode=itxm20
N2 - 3-dimensional (3D) cell cultures are being increasingly recognized as physiologically more relevantin vitromodels than traditional monolayer cultures, because they better mimicin vivo-like microenvironment, cell-cell and cell-extracellular matrix interactions. Nevertheless, the broader use of 3D models might be limited by requirements for special consumables, equipment, or skills for 3D cell cultures, and by their limited throughput and scalability. In this study, we optimized and adapted a commercially available agarose-micromolding technique to produce scaffold-free spheroid cultures. Brightfield microscopy was used for routine nondestructive and noninvasive evaluation of spheroid formation and growth. The workflow is compatible with manual, as well as high speed automated microscopic image acquisition, and it is supplemented with an in-house developed macro 'Spheroid_Finder' for open source software Fiji to facilitate rapid automated image analysis. This protocol was used to characterize and quantify spheroid formation and growth of two different hepatic cell lines, hTERT immortalized, but non-cancerous, adult human liver stem cell line HL1-hT1, and human hepatocellular carcinoma cell line HepG2, as well as their responses to a model antiproliferative and cytotoxic agent, 5-fluorouracil. The complete protocol provides a simple and ready-to-use solution to initiate scaffold-free spheroid cultures in any laboratory with standard equipment for mammalianin vitrocell culture work. Thus, it allows to increase throughput and scale of spheroid culture experiments, which can be greatly utilized in different areas of biomedical, pharmaceutical and toxicological research.
ER -
BASU, Amrita, Aneta DYDOWICZOVÁ, James E. TROSKO, Luděk BLÁHA a Pavel BABICA. Ready to go 3D? A semi-automated protocol for microwell spheroid arrays to increase scalability and throughput of 3D cell culture testing. \textit{Toxicology Mechanisms and Methods}. Taylor and Francis Ltd., 2020, roč.~30, č.~8, s.~590-604. ISSN~1537-6516. Dostupné z: https://dx.doi.org/10.1080/15376516.2020.1800881.
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