par Penasse, Willy 
;Bernaert, Denise ;Mosselmans, Roger
Référence PASCAL. F 52, Biochimie, biophysique moléculaire, biologie moléculaire et cellulaire, 34, 2-3, page (175-186)
Publication Publié, 1979
;Bernaert, Denise ;Mosselmans, Roger Référence PASCAL. F 52, Biochimie, biophysique moléculaire, biologie moléculaire et cellulaire, 34, 2-3, page (175-186)
Publication Publié, 1979
Article révisé par les pairs
| Résumé : | Cell surface properties of adult rat hepatocytes in situ, isolated by enzymatic perfusion and cultured have been examined by scanning electron microscopy. The parenchymal cells in situ, fixed by glutaraldehyde perfusion, reveal a well known surface heterogeneity: interhepatocytic faces exhibit villous bile canaliculi delineated by smooth surfaces and sinusoidal faces covered with numerous short and thick microvilli. During the isolation procedure, different steps in cell dissociation were distinguished, characterized by a progressive loss of cell membrane differentiations. Isolated hepatocytes present a homogeneous cell surface entirely covered with numerous microvilli. Although an important shrinkage (~ 30%) of the mean cell diameter of isolated hepatocytes was observed after critical point drying, their cell surface characteristics confirm previous data obtained by TEM. Different hepatocytic subpopulations were separated by elutriation, according to their density, size and ploidy. The counterflow centrifugation does not affect the cell surface ultrastructural properties. Liver parenchymal cells cultured in Dulbecco's medium supplemented with 17% fetal calf serum, aggregate and reconstitute cell trabeculae. Intimate cell contacts are established and newly formed bile canaliculi develop after 24 h at the level of the interhepatocytic faces. The latter are visualized in the SEM by artefactual fractures, which occur preferentially in the intercellular space, during critical point drying. Poly-L-lysine considerably enhances cell spreading and affects the distribution of microvilli. |
