An improved method for culturing patient-derived colorectal cancer spheroids

Oncotarget
Hiroyuki MiyoshiM Mark Taketo

Abstract

Recent advances allowed culturing and examination of patient-derived colorectal cancer (PD-CRC) cells as organoids or spheroids. To be applied to practical personalized medicine, however, current methods still need to be strengthened for higher efficiency. Here we report an improved method to propagate PD-CRC tumor initiating cells (TICs) in spheroid culture. We established > 100 cancer spheroid lines derived from independent colorectal cancer patients employing a serum-containing medium with additional inhibitors, Y27632 and SB431542. Because colorectal cancer spheroids showed wide-range growth rates depending on the patient tumors, we searched for supplementary factors that accelerated proliferation of slow-growing CRC-TIC spheroids. To this end, we introduced a convenient growth-monitoring method using a luciferase reporter. We found that epidermal growth factor (EGF) and/or basic fibroblast growth factor (bFGF) were critical for steady propagation of a subset of CRC-TIC spheroids carrying the wild-type RAS and RAF genes. We also identified 5'-(N-ethyl-carboxamido)-adenosine (NECA), an adenosine receptor agonist, as an essential supplement for another subset of spheroids. Based on these results, we propose to optimize cultur...Continue Reading

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Citations

Oct 18, 2018·Stem Cells Translational Medicine·Amar DesaiStanton L Gerson
Jun 4, 2020·Cancers·George M RamzyPatrycja Nowak-Sliwinska
Jun 20, 2019·ELife·Anthony EssexUNKNOWN Reproducibility Project: Cancer Biology
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Mar 6, 2021·Cancer Cell International·Se Jik HanKyung Sook Kim
Jun 27, 2020·Current Medicinal Chemistry·Soraia FernandesGiancarlo Forte

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Methods Mentioned

BETA
xenografts

Software Mentioned

Integrative Genomics Viewer
OLYMPUS
GraphPad PRISM
DP Controller
cellSens Standard
PRISM
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