Figure 4.

Functional Comparison of Surface and Crypt CD44⁺NGFR⁺ Cells

(A) Experimental protocol for studying the activities of FACS-purified CD44⁺NGFR⁺ cells in 2D CFC and organotypic 3D culture systems.

(B) Linear relationship between the number of cells seeded and the number of colonies formed per well in CFC assays.

(C) Hematoxylin staining of 14-day colonies generated in CFC assays.

(D) Number of CFCs detected per 100 input cells from each of the four subpopulations analyzed (R1, R2, R3, and R4, as shown in Figures 2D and 2E) in four experiments.

(E) Distribution of CFCs in each of the four surface and crypt subpopulations analyzed in four experiments.

(F and G) IHC staining of epithelial (Ep) tissue layers generated in organotypic 3D cultures of unsorted (top) and purified CD44⁺NGFR⁺ cells (bottom). Shown is the expression of markers specific for basal and/or parabasal cells (F) or differentiated cells (G). Arrowhead, Ki67⁺ cell. Scale bars, 100 μm.

See also Figure S1.