Interaction of liposomes with Kupffer cells in vitro
| Autor: | W.J.M. van Galen, Dharamdajal Kalicharan, Gerrit L. Scherphof, Caesar E. Hulstaert, Jan Kornelis Dijkstra, Frits H. Roerdink |
|---|---|
| Rok vydání: | 1984 |
| Předmět: |
Kupffer Cells
Antimycin A Horseradish peroxidase chemistry.chemical_compound Phosphatidylcholine Animals Bovine serum albumin Incubation Cells Cultured Liposome biology Vesicle Albumin Temperature Rats Inbred Strains Cell Biology Endocytosis Rats Kinetics Microscopy Electron Dextran Blood chemistry Biochemistry Microscopy Fluorescence Liposomes Vacuoles Biophysics biology.protein Sodium Fluoride Female Adsorption Lysosomes |
| Zdroj: | Experimental cell research. 150(1) |
| ISSN: | 0014-4827 |
| Popis: | We investigated the interaction of liposomes with rat Kupffer cells in monolayer maintenance culture. The liposomes (large unilamellar vesicles, LUV) were composed of 14 C-labelled phosphatidylcholine, cholesterol and phosphatidylserine (molar ratio 4 : 5 : 1) and contained either 3 H-labelled inulin or 125 I-labelled bovine serum albumin as a non-degradable or a degradable aqueous space marker, respectively. After 2–3 days in culture the cells exhibited optimal uptake capacity. The uptake process showed saturation kinetics, maximal uptake values amounting to 2 nmol of total liposomal lipid/h/106 cells. This is equivalent to 1500 vesicles per cell. The presence of fetal calf serum (FCS) during incubation increased uptake nearly two-fold, whereas freshly isolated rat serum had no effect. The binding of the liposomes to the cells caused partial release of liposomal contents (about 15–20%) both at 4°C and at 37°C. In the presence of metabolic inhibitors the uptake at 37°C was reduced to about 20% of the control values. Inulin and lipid label became cell-associated at similar rates and extents, whereas the association of albumin label gradually decreased after attaining a maximum at relatively low values. When, after 1 h incubation, the liposomes were removed continued incubation for another 2 h in absence of liposomes led to an approx. 30% release of cell-associated lipid label into the medium in water-soluble form. Under identical conditions as much as 90% of the cell-associated albumin label was released in acid-soluble form. Contrarily, the inulin label remained firmly cell-associated under these conditions. From these results we conclude that Kupffer cells in monolayer culture take up liposomes primarily by way of an adsorptive endocytic mechanism. This conclusion was confirmed by morphological observations on cells incubated with liposomes containing fluorescein isothiocyanate (FITC) dextran or horseradish peroxidase as markers for fluorescence microscopy and electron microscopy, respectively. |
| Databáze: | OpenAIRE |
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