Hyaluronic acid hydrogels with defined crosslink density for the efficient enrichment of breast cancer stem cells

Abstract

Cancer stem cells (CSCs) have been much proposed as potential tumor eradication targets since they possess highly tumorigenic qualities. However, efficient and fast enrichment of CSCs for cancer biology study and drug screening has been challenging. CD44 is a cell surface receptor for hyaluronic acid (HA) and has been reported as an important CSC marker. Here, we show a simple and label-free method for the enrichment of CSCs highly expressing CD44 using enzymatically crosslinked HA hydrogels. HA hydrogels were formed with different crosslink densities to modulate the interaction between the CD44 and HA chains. We show that HA hydrogels with defined crosslink densities isolated cancer cells expressing high CD44 from breast cancer cell lines in a facile, efficient manner. The enriched cells exhibited CSC-like characteristics such as high expression of CSC markers (octamer-binding transcription factor 4 (OCT4) and aldehyde dehydrogenase 1 (ALDH1)), enhanced tumorsphere formation and chemoresistance. The enriched cells also displayed strong tumorigenicity, metastatic potential and poor survival in vivo. The HA hydrogel provides a simple, fast and efficient platform for CSC enrichment and promotes new anticancer strategies that target breast CSCs. Statement of SignificanceThere is strong interest in developing isolation methods for cancer stem cells (CSCs), due in growing desire for CSC eradication for promising cancer therapy. Tumor sphere formation and fluorescence-activated cell sorting have been widely used for CSC isolation, while these methods require cultivation for several days and labelling of cell surface proteins, respectively. A simple and label-free method for breast CSC isolation is developed using HA-based hydrogels with tunable crosslink density. The efficient enrichment of breast CSCs is achieved by HA–CD44 specific interaction, which is controlled by hydrogel crosslink density. We believe that the simple approach that isolates cells with CSC-like characteristics would facilitate the anticancer drug development and cancer research.