Development of New Methods for the Detection, Isolation, Growth, and Re-collection of Cancer Cells In Blood Based on Size-discrimination of Cancer Cells and Normal Cells
2 Research Institute for Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan
4 Faculty of Industrial Scienc and Technology, Tokyo University of Science, 6-3-1 Nijuku, Katsushika-ku, Tokyo 125-8585, Japan
5 Laboratory of Veterinary Surgery, Joint Department of Veterinary Medicine, Faculty of Agriculture; Tottori University, Tottori 680-8553, Japan
Circulating tumor cells (CTCs) are tumor cells that are considered to originate from primary cancer sites, go into the blood stream in the body, and metastasize to the other organs. Herein, we report on a simple and convenient method to trap, culture, and recollect cancer cells, the sizes of which are greater than that of normal hematologic cells by the use of glass-beads filters (GBF) having a diameter of 24 mm and a thickness of 0.4~1.2 mm, which were prepared by sintering round-shaped glass beads (diameter: 63–106 m). A small integrated glass-beads filter (iGBF) with a diameter of ca. 9.6 mm for the use in filtering a small volume of blood was also designed and prepared. It was possible to efficiently capture mouse Lewis lung carcinoma cells expressing green fluorescent protein spiked in saline/blood by single and repeated (circulation) filtrations. In addition, we successfully captured B16 CTCs from the blood of a B16 melanoma metastasis mouse model by iGBF and grow them on/in iGBF. Filtration by GBF had negligible effect on the adherent and proliferative characteristics of cancer cells. We believe that the GBF protocols afford easy and efficient methods for early and convenient diagnosis and treatment of cancer and related diseases.
Keywords: Glass-bead Filter, Filtration, Circulating Tumor Cells, Size Based Separation