In VitroStudies on the Effect of Erythropoietin on Glucosamine-l-14C Incorporation Into Rat Bone Marrow Cells

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Abstract
It has been shown that glucosamine-l-14C is incorporated into the stroma (acid-insoluble fraction) of rat bone marrow cells in culture. Erythropoietin preparations at stages of purification ranging from untreated anemic plasma to one highly purified material (approximately 900 U/mg protein) caused increased glucosamine uptake into these bone marrow cell cultures. It is therefore assumed that erythropoietin is the agent responsible for this effect, which first becomes measurable after about 3 hr of incubation. A fixed amount of incorporating capacity appears after erythropoietin is added to the culture. The response to erythropoietin is a logarithmic function of the dose over a considerable part of the range tested. The radioactivity incorporated into the stroma has been identified in 3 classes of compounds: a) material not solubilized by 0.05N H2SO4 at 80 C (44% of total radioactivity in the original sample); b) material so far not identified which stayed at the origin of the paper-chromatographic system (21% of total radioactivity); c) material which appeared to be a mixture of N-acetyl-neuraminic acid and Nglycolyl-neuraminic acid (27 % of total radioactivity). (Endocrinology74: 960, 1964)