Simple Detection and Culture of Circulating Tumor Cells from Colorectal Cancer Patients Using Poly(2-Methoxyethyl Acrylate)-Coated Plates
Masatoshi Nomura, Yuhki Yokoyama, Daishi Yoshimura, Yasuhisa Minagawa, Aki Yamamoto, Yukiko Tanaka, Naoko Sekiguchi, Daiki Marukawa, Momoko Ichihara, Hiroaki Itakura, Kenichi Matsumoto, Yoshihiro Morimoto, Hideo Tomihara, Akira Inoue, Takayuki Ogino, Norikatsu Miyoshi, Hidekazu Takahashi, Hidenori Takahashi, Mamoru Uemura, Shogo Kobayashi, Tsunekazu Mizushima, Takahisa Anada, Masaki Mori, Yuichiro� Doki, Masaru Tanaka, Hidetoshi Eguchi, Hirofumi Yamamoto
Abstract
Here we aimed to establish a simple detection method for detecting circulating tumor cells (CTCs) in the blood sample of colorectal cancer (CRC) patients using poly(2-methoxyethyl acrylate) (PMEA)-coated plates. Adhesion test and spike test using CRC cell lines assured efficacy of PMEA coating. A total of 41 patients with pathological stage II–IV CRC were enrolled between January 2018 and September 2022. Blood samples were concentrated by centrifugation by the OncoQuick tube, and then incubated overnight on PMEA-coated chamber slides. The next day, cell culture and immunocytochemistry with anti-EpCAM antibody were performed. Adhesion tests revealed good attachment of CRCs to PMEA-coated plates. Spike tests indicated that ~75% of CRCs from a 10-mL blood sample were recovered on the slides. By cytological examination, CTCs were identified in 18/41 CRC cases (43.9%). In cell cultures, spheroid-like structures or tumor-cell clusters were found in 18/33 tested cases (54.5%). Overall, CTCs and/or growing circulating tumor cells were found in 23/41 CRC cases (56.0%). History of chemotherapy or radiation was significantly negatively correlated with CTC detection (p = 0.02). In summary, we successfully captured CTCs from CRC patients using the unique biomaterial PMEA. Cultured tumor cells will provide important and timely information regarding the molecular basis of CTCs.