Table 1.

Cancer stem cell quantification in xenografts by Fluorescence Activated Cell Sorting (FACS)

Pancreatic cancer xenografts	% ALDH+ cells	% CD44+CD24+ cells
Panc215	9.41	3.70
Panc265	0.82	8.47
Panc354	0.75	16.60
Panc266	4.67	0.64

NOTE: Four individual patient-derived pancreatic tumors grown subcutaneously (s.c.) in nude mice were harvested in sterile conditions. Single cell suspensions were generated by mincing tumors, followed by incubation in Dispase and Collagenase IV (Sigma) at 37°C for 2h.

The cell suspension was filtered and further purified by density centrifugation using Ficoll-Paque Plus (GE Healthcare). Cells were labeled using the ALDEFLUOR reagent (Stem Cell Technologies) and then stained with antibodies against mouse CD31 (BD Biosciences), lineage cocktail (Miltenyi Biotec), and H-2Kd (BD Biosciences). Non-mouse cells were separated and stained with antibodies against human CD44, CD24 (BD Biosciences). Cells were analyzed using FACSAria Flow Cytometer (BD Biosciences).