Retention of differentiated properties in an established dog kidney epithelial cell line (MDCK).

Abstract

Madin-Darby canine kidney (MDCK) cells grown in tissue culture have the morphological properties of distal tubular epithelial cells, form tight junctions, and lack several proximal tubular enzyme markers. Adenylate cyclase in these cells was stimulated by vasopressin, oxytocin, prostaglandins E1 and E2, glucagon, and cholera toxin. Hormone-stimulated adenylate cyclase activity in isolated membrane preparations was dependent on low concentrations of GTP and had the MgCIz and pH optima expected for the kidney enzyme. The results, as well as the demonstration of enhanced hemicyst formation induced by cyclic AMP, suggest that the MDCK cell line has retained the differentiated properties of the kidney epithelial cell of origin.When MDCK cells were injected into baby nude mice, continuous nodule growth was observed until adulthood was attained. Histological studies revealed the presence of two cell types: normal mouse fibroblasts which comprised 80-90% of the solid nodule mass, and MDCK cells, which formed epithelial sheets lining internal fluid-filled glands. Electron microscope analysis showed that the mucosal surfaces of the cells were characterized by microvilli which faced the lumen of the glands, that adjacent MDCK cells were joined by tight junctions, and that the serosal surfaces of the epithelial sheets were characterized by smooth plasma membranes which were lined by a continuous basement membrane. These observations lead to the conclusion that the MDCK cells retain regional differentiation of their plasma membranes and the ability to regenerate kidney tubule-like structures in vivo. KEY WORDS kidney epithelial cells epithelial cell polarization vasopressin prostaglandins E1 and E2cyclic AMP MDCK Processes of central importance to the renal physiologist include: (a) the biogenesis of functional kidney tubules, (b) transepithelial salt and fluid transport, and (c) hormonal regulation of transport functions (10, 34). Attempts to gain information about these processes have been hampered by the cellular complexity of the kidney. In general, it has not been possible to assign specific functions to individual cell types because pure populations of kidney-derived primary cell cul-J. CELL BIOLO6V 9 The Rockefeller University Press 9

Involvement of EP1 and EP2 receptors in the regulation of the Na,K-ATPase by prostaglandins in MDCK cells

“…Previous observations that PGE 1 and PGE 2 cause an increase in intracellular Ca 2+ and cAMP in MDCK cells are consistent with the involvement of EP1 and EP2 receptors in mediating the effects of these two prostaglandins in MDCK cells [10,16,19,30,[39][40][41][42]. The increase in intracellular Ca 2+ that occurs in PGE 1 and PGE 2 treated MDCK cells may cause a number of Ca 2+ mediated pathways to be activated (in addition to PKC), which may all contribute to the PGE 1 stimulation.…”

“…Indeed, arginine vasopressin, norepinephrine, glucagon, PGE 1 and PGE 2 all cause the activation of adenylate cyclase in MDCK cells [10,15,16]. In vitro studies concerning the actions of such effector molecules are facilitated by a hormonally defined culture conditions.…”

“…Adjacent cells are interconnected by tight junctions. At confluence, MDCK monolayers possess the capacity for transepithelial ion transport [10][11][12][13]. The Ussing chamber studies of Cereijido et al [14] have shown that (1) the entry of sodium into MDCK cells is dependent upon an amiloride-sensitive Na + /H + antiport system, and (2) the active extrusion of Na + in the apical to the basolateral direction is dependent upon a ouabain-sensitive Na,K-ATPase, which is localized on the basolateral surface [13].…”

Bovine colostrum supports the serum-free proliferation of epithelial cells but not of fibroblasts in long-term culture

“…MDCK is an established canine kidney epithelial cell line that preserves the structure and function of kidney epithelium (13,18) . The growth of MDCK cells in DMEM supplemented with various concentrations of milk and serum is shown in Fig .…”

The Exocyst Affects Protein Synthesis by Acting on the Translocation Machinery of the Endoplasmic Reticulum

“…Madin-Darby canine kidney (MDCK) cells represent one of the best studied polarized epithelial cell lines and are derived from canine renal tubular epithelium (8). When grown on a permeable filter, MDCK cells form a well polarized epithelial monolayer, exhibiting apical and basolateral plasma membrane domains with unique compositions (9,10), and well defined cell-cell junctional complexes containing tight junctions (11).…”