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Fig. 4. SP cells do not contribute to primary epithelial air liquid interface cultures. Primary epithelial cells were isolated from the tracheas of ROSA26 (A) and C57Bl/6 (B) mice and shown to form differentiated mature tracheal epithelium when 2x10⁵ cells were plated onto a transwell insert. The presence of ROSA26 cells was assessed by X-gal staining (A) of the cultures. However, when 2x10⁴ ROSA26 SP cells isolated from bone marrow were mixed with 2x10⁵ unmarked C57Bl/6 tracheal epithelial cells under the same culture conditions, they were not found to contribute to the formation of the tracheal epithelium (C) as assessed by lack of blue cells following staining with X-gal. These experiments were performed four times with three replicates in each experiment. The black dots that appear on the photomicrographs are transwell pores. Magnification x40.

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