Cytochemical identification of human granulo-monopoietic colonies grown in agar gel
Wang, S.Y.; Ho, C.K.; Chien, K.Y.
Journal of the Formosan Medical Association 86(3): 215-221
1987
ISSN/ISBN: 0371-7682 PMID: 3496422 Document Number: 287947
Human bone marrow hematopoietic cells were grown in soft agar in the presence of different sources of granulocyte-macrophage colony stimulating activity (GM-CSA). At days 7 and 14 post-incubation, the number and type of colonies were examined by a modified cytochemical methods involving the assay for esterase activities. The cells in agar gel were fixed in situ with four different fixatives under different conditions and then demonstrated for the presence of reactivity to .alpha.-naphthyl acetate (non-specific esterase, NSE) or naphthol AS-D chloroacetate (CAE) as specific enzyme markers for monocytoid and granulocytic cells respectively. Among the 4 different fixation methods used, cold (4.degree. C) 10% formol-calcium solution yielded the best result in preserving both colony morphology and esterase activity. The type of colonies formed at day 7 were mostly granulocytic, while more monocyte/macrophage colonies could be found at day 14. Furthermore, higher concentrations of GM-CSF favor the formation of granulocytic colonies and colonies induced by lower concentrations of GM-CSA were predominantly composed of monocytes and macrophages. Lastly, the proportion of granulocyte and macrophage mixed colonies remains fairly constant at all concentrations tested. The implications of these results are briefly discussed.