Association of calcium oxalate monohydrate crystals with MDCK cells.
Many factors are presently known which determine the risk of calcium oxalate (CaOx) stone formation in the kidney, although the early events in the pathogenesis of this disease are still to be elucidated. One of these early events is the interaction of intraluminal crystals with the epithelial cells lining the renal tubules. In this study we determined the interaction of approximately 2 microns calcium oxalate monohydrate (COM) crystal with monolayers of Madin-Darby canine kidney (MDCK) cells grown on porous supports in a two-compartment culture system. Crystal-cell interaction studies were performed after the monolayers reached their highest level of gamma-glutamyltranspeptidase (gamma GT) enzyme activity, a marker for brush border development. Technical aspects were evaluated, such as the size and morphology of the crystals and the influence of incubation time, temperature and pH on crystal-cell interaction. Kinetic data demonstrated that an equilibrium between free and associated particles was reached within 30 minutes. Crystal-cell interaction was often associated with cell damage. However, evidence is provided that in an environment that was saturated with calcium oxalate, MDCK cells in an environment that was a certain amount of COM crystals without sustaining measurable injury. After initial attachment to the cell surface, crystals were taken up and subsequently eliminated again from the monolayers. The model system described in this paper provides a tool for detailed studies of processes that are involved in renal cellular handling of luminal COM crystals.
Verkoelen
C F
CF
Department of Urology, Erasmus University Rotterdam, The Netherlands.
Romijn
J C
JC
de Bruijn
W C
WC
Boevé
E R
ER
Cao
L C
LC
Schröder
F H
FH
D000704:Analysis of Variance; D000818:Animals; D002129:Calcium Oxalate / Q000009:adverse effects / Q000378:metabolism*; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D003460:Crystallization; D004285:Dogs; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007685:Kidney Tubules, Collecting / Q000166:cytology / Q000378:metabolism*; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D013696:Temperature; D014961:X-Ray Diffraction; D005723:gamma-Glutamyltransferase / Q000378:metabolism
D002129:Calcium Oxalate; D007770:L-Lactate Dehydrogenase; D005723:gamma-Glutamyltransferase