We have been successful at isolating a "SP" population of epidermal cells (low Hoechst 33342 and PI content) that we called Epidermal Progenitor Cells. These cells are sensitive to Verapamil 50 uM. All these experiments were done with neonatal cells. We are now trying to isolate the same population of cells from adult skin. Our protocol calls for 90 min at 37 degrees in Hoechst, then resuspend in PI and sort. When we do that, we are able to select a population of cells that are verapamil sensitive. However, we have a huge percentage of dead cells within this population (80%). We have reduced the incubation time in the Hoechst to 30 min, and have seen a major improvement in the death rate (50%). However, after 30 min, they are not verapamil sensitive. The question is: can the verapamil be effective in 30 min, or not? Can the Hoechst be effective in 30 min or not? Thanks for helping us out. Martine -- -------------------------------- Martine Dunnwald, Pharm.D., Ph.D. Dermatology Research Department 2110A Med Lab The University of Iowa Iowa City (IA) 52242 tel:319-384-4645 fax:319-353-5615 martine-dunnwald@uiowa.edu http://tray.dermatology.uiowa.edu/M-Dunnwald.htmReceived on Thu Dec 16 13:18:00 2004
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