| Autor: |
McGinnes, Kimberley, Keystone, Edward, Bogoch, Earl, Hastings, David, Messner, Hans A., Jamal., Nazir, Paige, Christopher J. |
| Zdroj: |
Blood; September 1990, Vol. 76 Issue: 5 p896-905, 10p |
| Abstrakt: |
A reproducible method for growing normal human bone marrow B-lineage colonies in agar is described. The clono-genic cells require a rich medium, Opti-MEM (GIBCO/BRL, Burlington, Ontario, Canada), and a source of T-cell-derived factors for growth. The conditions described appear to be limiting for the colony progenitor, suggesting assay clonality. Three novel methods that permit routine and rapid detection of these human B-cell colonies are also described. Colonies containing cells that secrete immuno- globulin are detected by plaquing and protein immunoblot-ting, while RNA transcripts can be detected by RNA colony blotting. The detection of more than one secreted immunoglobulin isotype or RNA species in a single colony can also be achieved. This B-cell colony assay and the associated detection methods will allow the further delineation of human B lymphopoiesis in both normal and disease states. © 1990 by The American Society of Hematology. |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
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