Survival of human bone marrow progenitor cells after freezing: Improved detection in the colony-formation assay |
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Authors: | T Schlunk E Rüber M Schleyer |
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Institution: | Department of Clinical Physiology and Occupational Medicine, Protein Laboratory of the Department of Internal Medicine I, University of Ulm, Ulm, Germany |
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Abstract: | If cryopreserved suspensions of human bone marrow were stimulated by human placental conditioned medium in the same way as fresh unseparated marrows, less than 40% of granulopoietic progenitor cells (CFUc) was identified. By adding α-thioglycerol (0.6 mM) to the culture medium, the concentration of detectable CFUc in cryopreserved bone marrow was increased by a factor of 3.4, and the recovery of CFUc after cryopreservation rose to 90%. The low recovery of CFUc after freezing in the absence of α-thioglycerol is due to the destruction of accompanying cells. Noncolony-forming cells normally present in the fresh human marrow promote colony formation in cultures stimulated by placental conditioned medium. Their effect can be replaced by α-thioglycerol. It is concluded that, in order to detect all CFUc independent of the cellular composition of the marrow suspension, this supplement is essential for CFUc cultures stimulated by conditioned medium. |
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