Isolation and separation of toad bladder epithelial cells

Détails

ID Serval
serval:BIB_AC01B65987BA
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Isolation and separation of toad bladder epithelial cells
Périodique
Journal of Membrane Biology
Auteur(s)
Rossier  M., Rossier  B. C., Pfeiffer  J., Kraehenbuhl  J. P.
ISSN
0022-2631
Statut éditorial
Publié
Date de publication
07/1979
Peer-reviewed
Oui
Volume
48
Numéro
2
Pages
141-66
Notes
Journal Article --- Old month value: Jul 16
Résumé
The epithelium of the urinary bladder of Bufo marinus is composed of 5 cell types, i.e., granular (Gr), mitochondria-rich (MR) and goblet (G) cells which face the urinary lumen, microfilament-rich (MFR) and undifferentiated cells (Un) located basally. The epithelium was dissociated by collagenase and EGTA treatment. Fractionation of dispersed cells by isopycnic centrifugation on dense serum albumin solutions yielded 4 fractions: (i) a very light fraction (p approximately equal to 1.025) enriched in MR and MFR cells; (ii) a light fraction (p approximately equal to 1.045) enriched in vacuolated Gr cells; (iii) a heavy fraction (p approximately equal to 1.065) composed essentially of aggregated Gr cells, and (iv) a pellet (p approximately equal to 1.085) enriched in G and undifferentiated cells. Recoveries were based on cell counts and DNA measurements. DNA content per cell was 13.2 pg +/- 0.9 (n = 37). From 1 g fresh tissue, 62 +/- 5 x 10(6) (n = 10) cells were recovered before isopycnic centrifugation of which about 70% excluded Trypan blue. After centrifugation, 90 to 95% of the cells excluded the vital dye and approximately 3(9) x 10(6) cells were recovered from the gradient. Cell metabolism in each fraction was estimated by oxygen consumption measurements in absence or presence of ouabain, acetazolamide, and dinitrophenol. The consumption measurements in absence or presence of ouabain, acetazolamide, and dinitrophenol. The consumption was threefold higher in the very light and light fractions when compared to the heavy and pellet fractions. Ouabain sensitive oxygen consumption (QO2) represented 12 to 35% of the total O2 consumption depending on the cell fraction, and acetazolamide sensitive QO2 varied from -0.8% in the heavy fractions to 20% in the lighter fractions. DNP increased QO2 in all fractions by 20 to 50%. Finally, the cells were able to reaggregate and form junctional complexes upon addition of calcium to the medium.
Mots-clé
Animals Bufo marinus Cell Separation/methods Epithelial Cells Epithelium/metabolism/ultrastructure Female Kinetics Male Microscopy, Electron Oxygen Consumption Urinary Bladder/*cytology/metabolism/ultrastructure
Pubmed
Web of science
Création de la notice
24/01/2008 14:01
Dernière modification de la notice
20/08/2019 16:16
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