Isolation and Characterization of Potential Cancer Stem Cells from Solid Human Tumors—Potential Applications

Abstract

Cancer stem cells (CSCs) are a subpopulation of cells within a heterogeneous tumor that have enhanced biologic properties e.g., increased capacity for self-renewal, increased tumorigenicity, enhanced differentiation capacity, and resistance to chemo-and radiotherapies. This unit describes protocols to isolate and characterize potential cancer stem cells from a solid tumor. This involves creating a single-cell suspension from tumor tissue, tagging the cell subpopulation of interest, and sorting cells into different populations. Finally, the sorted subpopulations can be evaluated for their ability to meet the functional requirements of a CSC, which primarily include increased tumorigenicity in an in vivo xenograft assay. Mastering the protocols in this unit will allow the researcher to study populations of cells that may have properties of CSCs.

Ovarian cancer stem cells: Are they real and why are they important?

“…Multiple experimental models have been used to potentially isolate putative cancer stem cells (Table 1). (19) Bapat and colleagues were one of the first groups to demonstrate heterogeneous growth properties in ovarian cancer cells in 2005 (20). They isolated unsorted ascitic cells from a patient and developed 19 spontaneously immortalized clones through low-density culturing.…”

ABC transporters in CSCs membranes as a novel target for treating tumor relapse

“…The detection can be performed with specific antibodies in flow cytometry, competitive ELISA (cELISA) or magnetic beads (Dobbin and Landen, 2013). Xia and coworkers have developed humanized antibodies toward the surface marker CD133 that was detected by cELISA (Xia et al, 2013).…”

Isolation and Characterization of Stem Cells from Human Central Nervous System Malignancies

“…Drugs such as verapamil, reserpine, or fumitremorgin C inhibit Hoechst exclusion and abolish the SP. Co-staining with antibodies helps with the identifi cation and confi rmation of the cells [ 11 ].…”

“…The most popular model is the NOD-SCID mouse model, which has a completely knocked out immune system [ 3 ]. However this model is susceptible to developing spontaneous lymphomas as early as within a year of life [ 11 ]. Other mouse models include IL2 receptor-γ chain defi cient, BALB/c-nude and Scid/bg [ 90 , 91 ].…”

“…For neurospheres, Singh and colleagues were the fi rst group to use an in vivo limiting dilution assay to implant fewer and fewer TSCs at each subsequent passage to assess the fewest number of cells required to form a tumor in the animal [ 22 ]. As a general principle, compared to the marker-negative population, at least 50-to 100-fold fewer marker-positive cells should be needed to stimulate tumor formation in 50 % of the mice [ 11 ] The mouse implantation model has some limitations as well. First, it is not possible to accurately assess the proportion of TSCs in original tumor mass.…”