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Isolation, propagation and characterization of primary tubule cell culture from human kidney (Methods in Renal Research)
Author(s) -
QI WEIER,
JOHNSON DAVID W,
VESEY DAVID A,
POLLOCK CAROL A,
CHEN XINMING
Publication year - 2007
Publication title -
nephrology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.752
H-Index - 61
eISSN - 1440-1797
pISSN - 1320-5358
DOI - 10.1111/j.1440-1797.2007.00779.x
Subject(s) - percoll , proximal tubule , cytokeratin , in vitro , cell culture , kidney , microbiology and biotechnology , pathology , medicine , brush border , human kidney , biology , biochemistry , immunohistochemistry , vesicle , genetics , membrane
SUMMARY: Proximal tubule cells (PTC) are the major cell type in the cortical tubulointerstitium. Because PTC play a central role in tubulointerstitial pathophysiology, it is essential to prepare pure PTC from kidney tissue to explore the mechanisms of tubulointerstitial pathology. The authors have successfully refined and characterized primary cultures of human PTC using Percoll density gradient centrifugation as a key PTC enrichment step. The cells obtained by this method retain morphological and functional properties of PTC and are minimally contaminated by other renal cells. In particular, the primary isolates have characteristics of epithelial cells with uniform polarized morphology, tight junction and well‐formed apical microvilli. Cytokeratin is uniformly and strongly expressed in the isolates. Brush border enzyme activities and PTC transport properties are retained in the isolates. This method therefore provides an excellent in vitro model for the physiologic study of the human proximal tubule.