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Evaluation of a short term in vitro test for granulocyte chalone activity

Virchows Archiv B Cell Pathology 20(3): 229-238
Evaluation of a short term in vitro test for granulocyte chalone activity
A short term in vitro test for granulocyte chalone activity was examined for its specificity and reliability. The test uses the inhibition, by granulocyte extracts, of 3H-thymidine (3H-Tdr) uptake into the acid-insoluble material by rat bone marrow cells in vitro to measure possible chalone activity. Among the many possible 3H-Tdr artifacts pool size dilution by Tdr contained in the extracts was excluded using an Escherichia coli mutant requiring thymine. Several amino acids and biogenic amines do not affect the test. Continuous and pulse labeling of bone marrow cells with 3H-Tdr, viability tests and micro flow fluorometric measurements of the cell cycle distribution following Colcemid treatment strongly suggest that the cells do not proliferate in vitro during short term incubation, since practically no cells enter the S-phase, cells in the S-phase die and few if any cells proceed through G2 and mitosis. The test cannot exclude cytotoxicity. Thus, the in vitro test may often screen for an unspecific S-phase inhibitor and must hence be supplemented by another assay to prove the chalone nature of an extract or fraction. The test per se fails to meet most of the requirements of a valid granulocyte chalone assay.

Accession: 005395697

PMID: 132006

DOI: 10.1007/bf02890342

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