Ferritin, added to the incubation medium of ascites tumor cells, was used as an electron microscopic marker to study the uptake of large protein molecules by morphologically intact cells. A definite uptake could be detected after 1 hour of incubation in Tyrode bicarbonate solution containing 0.04 to 13.3 mg ferritin/ml. Ferritin was found in a variety of membrane-surrounded structures, suggesting that pinocytesis and related membrane movements are occurring under physiological conditions and can account for the penetration of intact macromolecules into isolated tumor cells. Supplementation of the medium with serum albumin (33 mg/ml) increased the average amount of ferritin per cell and per pinocytotic structure. Ferritin was strongly adsorbed by fragments of lysed cells, which were readily taken up by intact cells. Besides its role as carrier, this debris appeared to stimulate membrane movements. Only rare examples were found to suggest the release of ferritin from the pinocytotic structures into the cytoplasm. Thus, the disintegration of such structures cannot be considered an obvious step towards a rapid metabolic utilization of protein by the cell. Particles of colloidal gold presented to the cell under the same conditions were not taken up to any significant extent, thus providing good evidence for a selective ingestion of particles of comparable sizes.

Animals↗

PubMed1962

Studies on protein uptake by isolated tumor cells. II. Quantitative data on the adsorption and uptake of I-131-serum albumin by Ehrlich ascites tumor cells.

Surface adsorption is studied in some detail because it is believed to be a major artifact in measurements of protein uptake by mammalian cells. Adsorption increases linearly with the I(131)-albumin concentration between 0.001 and 300 mg/ml. After short exposure to 300 mg/ml and two cell washings, the adsorption amounts to 38 mg albumin per gm cell proteins. Further washings remove 80 per cent of this value, leaving a small irreversibly bound residue. At equilibrium, adsorbed albumin can be labeled by a simple albumin exchange. This labeling reaches a steady state within seconds and stays at constant level over 30 minutes. Significant increases above this initial level are measured over periods of 2 hours. In our experimental conditions these increases can be considered due to albumin uptake. This uptake rises linearly with the albumin concentration between 0.5 and 50.0 mg/ml, and reaches 0.2 mg/gm cell protein or 4 x 10(5) molecules per cell. Compared to the incorporation of free amino acids in similar conditions, this value does not appear to contribute significantly to the N-metabolism of the tumor cells. Adsorption was generally greater than uptake. Both processes are linear functions of the same variable over the whole range of concentration tested. It is suggested that albumin is taken up by pinocytosis.

Adsorption↗

PubMed1962