Background/Aim: Retention of crystals in the kidney ultimately leads to renal stone formation. Hyaluronan (HA) has been identified as binding molecule for calcium oxalate monohydrate crystals. The association of high molecular mass (M<sub>r</sub>) HA with cell surface receptors such as CD44 gives rise to pericellular matrix (PCM) formation by many eukaryotic cells in culture. Here, we study the ability of several renal tubular cell lines to assemble PCMs and to synthesize high-M<sub>r</sub> HA during proliferation in relation to crystal retention. Methods: PCM assembly by MDCK-I, MDCK-II, and LLC-PK<sub>1</sub> cells was visualized by particle exclusion assay. Metabolic labeling studies were performed to estimate the cellular production of HA. The expression of CD44 and HA was studied using fluorescent probes, and crystal binding was quantified with radiolabeled calcium oxalate monohydrate. Results: PCMs were formed, and HA was expressed by most MDCK-I and some MDCK-II, but not by LLC-PK<sub>1</sub> cells. All cell types expressed CD44 at their apical surface. MDCK-I and MDCK-II cells secreted, respectively, 14.7 ± 1.6 and 0.5 ± 0.2 pmol [<sup>3</sup>H]glucosamine incorporated in high-M<sub>r</sub> HA, whereas LLC-PK<sub>1</sub> cells did not secrete HA. Streptomyces hyaluronidase treatment significantly decreased crystal binding (µg/cm<sup>2</sup>) to MDCK-I cells (from 8.6 ± 0.4 to 3.9 ± 0.9), but hardly to MDCK-II cells (from 10.2 ± 0.2 to 9.6 ± 0.1) or LLC-PK<sub>1</sub> cells (from 10.2 ± 0.8 to 9.9 ± 0.3). Conclusions: There are various forms of crystal binding to renal tubular cells in culture. Crystal attachment to MDCK-I and some MDCK-II cells involves PCM assembly that requires high-M<sub>r</sub> HA synthesis. HA production and PCM formation do not play a role in crystal binding to LLC-PK<sub>1</sub> and the majority of MDCK-II cells. It remains to be determined which form of binding is involved in renal stone disease.