TY - JOUR
T1 - Multiparameter analyses of three-dimensionally cultured tumor spheroids based on respiratory activity and comprehensive gene expression profiles
AU - Zhou, Yuanshu
AU - Arai, Toshiharu
AU - Horiguchi, Yoshiko
AU - Ino, Kosuke
AU - Matsue, Tomokazu
AU - Shiku, Hitoshi
N1 - Funding Information:
This research was partly supported by the Cabinet Office, Government of Japan, through its “Funding Program for Next Generation World-Leading Researchers” (to H.S.).
PY - 2013
Y1 - 2013
N2 - Multicellular spheroids of human breast cancer cells (MCF-7) formed with two different three-dimensional (3D) culture methods were evaluated in detail on the basis of respiratory activity and highthroughput gene expression analysis. The spheroids formed with poly(dimethylsiloxane) (PDMS) microwell arrays indicated significant restriction of the spheroid size, whereas their respiratory activity was 2-fold greater than that formed with the hanging drop culture method. Fluidigm BioMark dynamic array was used for comprehensive and quantitative real-time polymerase chain reaction (qRT-PCR) analysis on the samples whose respiratory activity had been measured. Genes involved in cellular senescence and glucose metabolism indicated significantly higher values for the PDMS microwell culture method than for the hanging drop culture method (P < 0.05). Interestingly, samples formed with the PDMS microwell culture method showed stronger responses for glycolysis than those formed with the hanging drop method. These results illustrate the power of multiparameter analysis to characterize multicellular spheroids cultured in different microenvironments even if they have the same morphology.
AB - Multicellular spheroids of human breast cancer cells (MCF-7) formed with two different three-dimensional (3D) culture methods were evaluated in detail on the basis of respiratory activity and highthroughput gene expression analysis. The spheroids formed with poly(dimethylsiloxane) (PDMS) microwell arrays indicated significant restriction of the spheroid size, whereas their respiratory activity was 2-fold greater than that formed with the hanging drop culture method. Fluidigm BioMark dynamic array was used for comprehensive and quantitative real-time polymerase chain reaction (qRT-PCR) analysis on the samples whose respiratory activity had been measured. Genes involved in cellular senescence and glucose metabolism indicated significantly higher values for the PDMS microwell culture method than for the hanging drop culture method (P < 0.05). Interestingly, samples formed with the PDMS microwell culture method showed stronger responses for glycolysis than those formed with the hanging drop method. These results illustrate the power of multiparameter analysis to characterize multicellular spheroids cultured in different microenvironments even if they have the same morphology.
KW - High-throughput qPCR
KW - Multicellular tumor spheroids
KW - PDMS microwell array
KW - Respiratory activity
UR - http://www.scopus.com/inward/record.url?scp=84879213723&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84879213723&partnerID=8YFLogxK
U2 - 10.1016/j.ab.2013.04.020
DO - 10.1016/j.ab.2013.04.020
M3 - Article
C2 - 23628321
AN - SCOPUS:84879213723
SN - 0003-2697
VL - 439
SP - 187
EP - 193
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -
