Ready to go 3D? A semi-automated protocol for microwell
spheroid arrays to increase scalability and throughput of 3D
cell culture testing

J 2020

Ready to go 3D? A semi-automated protocol for microwell spheroid arrays to increase scalability and throughput of 3D cell culture testing

BASU, Amrita, Aneta DYDOWICZOVÁ, James E. TROSKO, Luděk BLÁHA, Pavel BABICA et. al.

Základní údaje

Originální název

Ready to go 3D? A semi-automated protocol for microwell spheroid arrays to increase scalability and throughput of 3D cell culture testing

Autoři

BASU, Amrita (356 Indie, domácí), Aneta DYDOWICZOVÁ (203 Česká republika, domácí), James E. TROSKO (840 Spojené státy), Luděk BLÁHA (203 Česká republika, domácí) a Pavel BABICA (203 Česká republika, garant, domácí)

Vydání

Toxicology Mechanisms and Methods, Taylor and Francis Ltd. 2020, 1537-6516

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

30108 Toxicology

Stát vydavatele

Velká Británie a Severní Irsko

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 2.987

Kód RIV

RIV/00216224:14310/20:00114569

Organizační jednotka

Přírodovědecká fakulta

DOI

http://dx.doi.org/10.1080/15376516.2020.1800881

UT WoS

000562645500001

Klíčová slova anglicky

Multicellular spheroids; 3D cell cultures; hepatotoxicity; in vitro toxicity testing

Štítky

rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 29. 4. 2021 12:35, Mgr. Marie Šípková, DiS.

Anotace

V originále

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.

Návaznosti

GA15-12408S, projekt VaV

Název: Role kmenových a diferencovaných jaterních buněk v hepatotoxicitě a hepatokarcinogenitě indukované cyanotoxiny

Investor: Grantová agentura ČR, Role kmenových a diferencovaných jaterních buněk v hepatotoxicitě a hepatokarcinogenitě indukované cyanotoxiny

GA19-19143S, projekt VaV

Název: 3-dimenzionální in vitro modely jater pro studium chronické hepatotoxicity a jaterních onemocnění indukovaných sinicovými toxiny.

Investor: Grantová agentura ČR, Jaterní 3D modely pro studium chronické hepatotoxicity a jaterních onemocnění indukovaných sinicovými toxiny

LM2018121, projekt VaV

Název: Výzkumná infrastruktura RECETOX (Akronym: RECETOX RI)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, RECETOX RI

Citovat

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. 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.

@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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