Genome and Transcriptome Profiles of CD133-Positive Colorectal Cancer Cells

A, B: Control group. From eight FFPE blocks, two needle biopsies of approximately 0.5-cm distance in either the CD133+ or CD133− tumor area were cut and analyzed with aCGH for genetic differences (E). After immunohistochemistry for CD133, (C) the CD133+ and CD133− cells were laser microdissected (D) and analyzed with array comparative genomic hybridization for genetic differences (E).

Staining was performed according to a conventional set-up used for primary CRC cells in the literature.25,26

The proportion of CD133+/CD133− in HCT-116 cultures was not significantly different to the set-up in Supplemental Figure 2. The staining procedure allowed identifying small changes in the CD133 distribution throughout culturing. In short-term cultures after separation, the populations preserved their CD133 expression profile. At later stages, we observed a redistribution of the CD133+ and CD133− cell populations in the cultured CD133+ fraction. Given are the number of passages (P) and of cumulative population doublings (CPD) after sorting. The culture doubling time for the two fractions was identical.

CD133+/CD133− distributions after tumor growth in xenografts derived from injection of either CD133+ or CD133− subpopulation estimated by FACS (A) after HT−29 cell injection and (B) after HCT 116 cell injection.