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Title: An improved method for culturing patient-derived colorectal cancer spheroids
Authors: Miyoshi, Hiroyuki
Maekawa, Hisatsugu
Kakizaki, Fumihiko  kyouindb  KAKEN_id
Yamaura, Tadayoshi
Kawada, Kenji
Sakai, Yoshiharu
Taketo, M. Mark
Author's alias: 三好, 弘之
前川, 久継
柿崎, 文彦
山浦, 忠能
河田, 健二
坂井, 義治
武藤, 誠
Keywords: colorectal cancer
chemical screening
cancer stem cell
tumor-initiating cell
Issue Date: 24-Apr-2018
Publisher: Impact Journals, LLC
Journal title: Oncotarget
Volume: 9
Issue: 31
Start page: 21950
End page: 21964
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 culture conditions for CRC-TIC spheroids by adjusting to the respective tumor samples. Our method provides a versatile tool that can be applied to personalized chemotherapy evaluation in prospective clinical studies.
Description: 患者由来大腸がん幹細胞培養を用いた薬剤感受性試験を開発 --個別化医療の実現へ期待--. 京都大学プレスリリース. 2018-09-03.
Rights: All site content, except where otherwise noted, is licensed under a Creative Commons Attribution 3.0 License.
DOI(Published Version): 10.18632/oncotarget.25134
PubMed ID: 29774115
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