Using of keratin substrate for enrichment of HT29 colorectal cancer stem‐like cells

Abstract Eradication of cancer stem‐like cells (CSLCs) are becoming increasingly an important target for new cancer therapies. The ability to study their behavior in vitro will provide the opportunity for high‐throughput testing of more effective treatments. In this study, spheroid‐like structures’ formation and enrichment of HT29 CSLCs were evaluated on a wool keratin‐based substrate as a bio‐mimic of natural extracellular matrix (ECM) proteins. The results indicated that culturing on keratin substrate increased spheroid formation ability and radio‐/chemoresistance of HT29 cells. Moreover, cell surface expression of CD133 CSLCs’ marker and the mRNA level of stemness genes such as Nanog, Oct4, and c‐MYC were increased. These data suggest that keratin can potentially be used for spheroid‐like structure formation and enrichment of HT29 CSLCs. In addition, it seems that the induction of stemness characteristics on keratin substrate is probably because of the activation of α 2 β 1 integrin signaling pathway. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1264–1271, 2019.