Isolation of Cancer Stem Cells by Side Population Method

Abstract

The Hoechst side population (SP) method is a flow cytometry technique used to obtain stem cells based on the dye efflux properties of the ATP-binding cassette (ABC) transporters. The SP cells are characterized by their capability to efflux the fluorescent DNA-binding dye Hoechst 33342 through their ABC transporters and are enriched in stem cells, which are endowed with a self-renewal capacity and multilineage differentiation potential and express the stemness genes including ABC multidrug transporters. The protocols outlined in this book chapter describe the isolation method of the SP cells from human lung carcinoma cell lines by using Hoechst 33342. In addition, we refer to the propagation method of SP cells by successive rounds of fluorescence-activated cell sorting analysis for SP cells. These approaches will be helpful for the establishment of novel in vitro and in vivo models using cancer stem cells, which may play a key role during carcinogenesis and/or tumor progression.

The Interactions Between Cancer Stem Cells and the Innate Interferon Signaling Pathway

“…IFN-α has also been reported to specifically target the side population (SP) of ovarian cancer cells, a subset of cells endowed with stem-like properties (i.e., CSCs) (158). In an attempt to exploit the anti-tumor effects of IFN-α, ovarian PDXs were subjected to gene therapy with IFN-α, and results showed a marked increase in survival rate in those PDXs bearing a high proportion of SP cells compared to those containing a low proportion, indicating that IFN-α specifically and negatively affects the CSC compartment.…”

Targeting Liver Cancer Stem Cells: An Alternative Therapeutic Approach for Liver Cancer

“…Stem cells with relatively high expression of ATP-binding cassette (ABC) transporter superfamily members show resistance against the absorption of unrelated (toxic) substances by pumping these compounds across cell membranes [ 108 ]. This action results in a low Hoechst 33342 side population (SP) [ 109 ]. In 2006, Chiba et al successfully used Hoechst dye 33342-effluxing SP cells to identify hepatic cancer cells with stem-like properties in HCC [ 110 ].…”

A low MW inhibitor of CD44 dimerization for the treatment of glioblastoma

“…GSCs can be enriched by the stem cell markers CD133 and CD44(Anido et al, 2010; Pietras, 2011; Singh et al, 2003). GSCs are also able to exclude Hoechst 33342 in the side population and are related to chemotherapy tolerance, as are other cancer stem cells (Shimoda, Ota, & Okada, 2018; Was et al, 2018).…”