Abstract
Human placental conditioned medium is able to stimulate human bone marrow cells to form neutrophilic granulocyte-monocyte colonies and eosinophil colonies in agar cultures. Methods are described for preparing highly active preparations (capable of supramaximal stimulation of colony formation) by partial purification of the conditioned medium using calcium phosphate absorption and gel filtration chromatography. Partially purified calcium phosphate gel eluates appear to increase in specific activity after concentration by ultrafiltration. This phenomenon was not observed with the active fractions after further purification on Sephadex G-100. Fractionation of the conditioned medium using gel filtration and hydrophobic chromatography (Cibacron Blue-Sepharose) showed that distinct and partially separable factors were responsible for stimulating granulocyte-macrophage and eosinophil colony formation. From the gel filtration data the apparent molecular weight of the eosinophil colony stimulating factor was higher than the molecular weight of the granulocyte-macrophage colony stimulating factor. Cibacron Blue-Sepharose chromatography led to the separation of one molecular species of granulocyte-macrophage colony stimulating factor from eosinophil colony stimulating factor so that a specific stimulus for human neutrophilic granulocyte and/or monocyte-macrophage progenitors is now available.
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