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Cell type-specific acquired protection from crystal adherence by renal tubule cells in culture
C F Verkoelen1, B G van der Boom, D J Kok
1Department of Urology, Erasmus University, Rotterdam, The Netherlands. Verkoelen@uro.fgg.eur.nl
Background:
Adherence of crystals to the surface of renal tubule epithelial cells is considered an important step in the development of nephrolithiasis. Previously, we demonstrated that functional monolayers formed by the renal tubule cell line, Madin-Darby canine kidney (MDCK), acquire protection against the adherence of calcium oxalate monohydrate crystals. We now examined whether this property is cell type specific. The susceptibility of the cells to crystal binding was further studied under different culture conditions.
Methods:
Cell-type specificity and the influence of the growth substrate was tested by comparing calcium oxalate monohydrate crystal binding to LLC-PK1 cells and to two MDCK strains cultured on either permeable or impermeable supports. These cell lines are representative for the renal proximal tubule (LLC-PK1) and distal tubule/collecting duct (MDCK) segments of the nephron, in which crystals are expected to be absent and present, respectively.
Results:
Whereas relatively large amounts of crystals adhered to subconfluent MDCK cultures, the level of crystal binding to confluent monolayers was reduced for both MDCK strains. On permeable supports, MDCK cells not only obtained a higher level of morphological differentiation, but also acquired a higher degree of protection than on impermeable surfaces. Crystals avidly adhered to LLC-PK1 cells, irrespective of their developmental stage or growth substrate used.
Conclusions:
These results show that the prevention of crystal binding is cell type specific and expressed only by differentiated MDCK cells. The anti-adherence properties acquired by MDCK cells may mirror a specific functional characteristic of its in situ equivalent, the renal distal tubule/collecting ducts.
Insights
Differentiated Madin-Darby canine kidney (MDCK) cells, representing the renal distal tubule, prevent calcium oxalate crystal adherence. This protective property is cell-type specific and not observed in LLC-PK1 cells, which model the renal proximal tubule.
Area of Science:
- Nephrology
- Cell Biology
- Biochemistry
Background:
- Crystal adherence to renal tubule epithelial cells is a key factor in nephrolithiasis development.
- Madin-Darby canine kidney (MDCK) cells, a renal tubule cell line, previously showed protection against calcium oxalate monohydrate crystal adherence.
- This study investigates if this protective property is cell-type specific.
Purpose of the Study:
- To determine if the anti-adherence property against calcium oxalate monohydrate crystals is specific to certain renal cell types.
- To investigate the influence of culture conditions and growth substrates on crystal binding to renal cells.
Main Methods:
- Compared calcium oxalate monohydrate crystal binding to LLC-PK1 (renal proximal tubule) and two MDCK (renal distal tubule/collecting duct) cell strains.
- Cultured cells on both permeable and impermeable supports to assess growth substrate influence.
- Evaluated crystal adherence under varying cell culture conditions and developmental stages.
Main Results:
- LLC-PK1 cells avidly bound crystals regardless of culture conditions.
- MDCK cells showed reduced crystal binding when confluent, especially on permeable supports.
- MDCK cells on permeable supports exhibited enhanced differentiation and greater protection against crystal adherence.
Conclusions:
- The prevention of crystal binding is a cell-type specific characteristic of differentiated MDCK cells.
- This anti-adherence property in MDCK cells may reflect a functional characteristic of the in situ renal distal tubule and collecting ducts.
- The findings suggest a potential mechanism for preventing kidney stone formation in specific nephron segments.