Conny wrote: >Hi, > >I am working with a FACS Vantage with TS option in a hematology/oncology >lab. We are sorting stem cell subpopulations in human leukapharesis >products. Some of the cells are frozen in plasma and 10 % DMSO like >normal blood cells for stem cell transplantation. After thawing in warm >water I quickly dilute the cells in PBS with 5mM EDTA and 10 %FCS and >wash them. But within a short time (about half an hour) cells will >agregate so that staining and sorting is impossible. Has anyone >suggestions how to avoid this clumping? I have tried trypsin and nylon >mesh to disconnect the cell masses, but that was not sucessful. Any >proposal is highly welcome since we have deepfreezed many interesting >samples from leukemia patients. > >Thanks in advance, > >Conny Brendel You might try using some DNAse in your media as the clumping may be caused by DNA from cells that have been destroyed through manipulations. Roger A. Burger E-mail: Roger@cpd2.usu.edu Research Associate, Immunology Center for Persons with Disabilities Utah State University Logan, UT 84322-6895 Voice: 801-797-2042 FAX: 801-797-4054
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