Print Email Facebook Twitter Isolation and characterization of Kupffer and endothelial cells from the rat liver Title Isolation and characterization of Kupffer and endothelial cells from the rat liver Author Instituut voor experimentele gerontologie TNO Knook, D.L. Blansjaar, N. Sleyster, E.C. Publication year 1977 Abstract Non-parenchymal cell suspensions were prepared from rat livers by three different methods based on a collagenase, a pronase and a combined collagenase-pronase treatment. The highest yield of Kupffer and endothelial cells was obtained with the pronase treatment. Attempts were made for a further purification of these cells by Metrizamide density gradient centrifugation after preferentially loading lysosomal structures in Kupffer cells with Triton WR 1339, Jectofer®, Neosilvol®, Zymosan or colloidal carbon. After loading with Triton WR 1339 or Jectofer®, highly purified endothelial cell suspensions were obtained, but the final Kupffer cell preparations were contaminated with about 20% of endothelial cells. Kupffer and endothelial cells purified in this way showed an altered ultrastructure and contained increased activities of the lysosomal enzymes acid phosphatase, arylsulphatase B and cathepsin D. As an alternative procedure for the purification of Kupffer and endothelial cells, a method based on centrifugal elutriation was employed. With this procedure, highly purified preparations of Kupffer or endothelial cells with a well preserved ultrastructure were obtained. Compared with endothelial cells, purified Kupffer cells had a three times higher cathepsin D activity, whereas the arylsulphatase B activity was three times higher in endothelial cells. The high cathepsin D activity in Kupffer cells could be nearly completely inhibited by the specific cathepsin D inhibitor pepstatin, which excludes a possible contribution to this activity by proteases endocytosed during the isolation of the cells. Chemicals/CAS: Cathepsins, EC 3.4.-; Chondro-4-Sulfatase, EC 220.127.116.11; Pronase, EC 3.4.24. Subject Endothelium cellAnimalCathepsinsCell SeparationCentrifugationCentrifugation, Density GradientChondro-4-SulfataseEndotheliumKupffer CellsLiverPronaseRats To reference this document use: http://resolver.tudelft.nl/uuid:4d94211f-3e24-49f4-958c-64b66ecd58f6 DOI https://doi.org/10.1016/0014-4827(77)90011-8 TNO identifier 228269 Source Experimental Cell Research, 109 (109), 317-329 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.