RRC ID 60022
Author Sato Y, Yamada T, Hiroyama T, Sudo K, Hasegawa N, Hyodo I, Nakamura Y.
Title A robust culture method for maintaining tumorigenic cancer stem cells in the hepatocellular carcinoma cell line Li-7.
Journal Cancer Sci
Abstract Cancer tissues contain small populations of highly tumorigenic cells termed cancer stem cells (CSCs). Immortalized cell lines containing CSCs are valuable and powerful experimental tools for research into the characteristics of these stem cells. We previously reported that the hepatocellular carcinoma cell line Li-7 includes abundant CD13+ CD166- CSCs; however, the number of these cells decreases after long-term culture as a result of differentiation to non-CSC populations. To ensure consistent and reproducible results in experiments using Li-7 cells, it is important that the CSC population is maintained stably regardless of culture duration and passage. In the present study, we found that a commercially available culture medium for maintenance of embryonic stem cells and induced pluripotent stem cells, mTeSR1, effectively prevented spontaneous differentiation by CD13+ CD166- cells to CD13- CD166+ cells and therefore maintained the CSC population in Li-7 cell cultures. CD13+ CD166- CSCs maintained using this culture medium retained high tumorigenicity after transplantation into mice; they also showed the ability to differentiate in vitro into non-CSC populations in RPMI-1640 with 10% FBS medium. We analyzed gene expression profiles of CSC and non-CSC populations in Li-7 cultures using an RNA sequencing method. Genes such as FGFR, NOTCH1, and JAG1, that are associated with tumorigenicity and stemness, were upregulated in the CSC population. Our results suggest that CSCs can be maintained in immortalized cancer cell lines cultured over an extended period using a medium developed for culture of embryonic/induced pluripotent stem cells.
Volume 110(5)
Pages 1644-1652
Published 2019-5-1
DOI 10.1111/cas.13978
PMID 30784169
PMC PMC6500967
MeSH Animals Antigens, CD / metabolism Biomarkers, Tumor / genetics* CD13 Antigens / metabolism Carcinoma, Hepatocellular / genetics Carcinoma, Hepatocellular / metabolism* Carcinoma, Hepatocellular / pathology Cell Adhesion Molecules, Neuronal / metabolism Cell Culture Techniques / methods* Cell Differentiation Cell Line, Tumor Cell Proliferation Culture Media / pharmacology Fetal Proteins / metabolism Gene Expression Profiling Humans Jagged-1 Protein / genetics Liver Neoplasms / genetics Liver Neoplasms / metabolism* Liver Neoplasms / pathology Mice Neoplasm Transplantation Neoplastic Stem Cells / cytology* Neoplastic Stem Cells / metabolism Neoplastic Stem Cells / transplantation* Receptor, Notch1 / genetics Receptors, Fibroblast Growth Factor / genetics Sequence Analysis, RNA Up-Regulation
IF 4.751
Times Cited 0
Human and Animal Cells Li-7(RCB1941)