Isolation, propagation and characterization of primary tubule cell culture from human kidney.
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.
Qi
Weier
W
Department of Medicine, University of Sydney, Kolling Institute, Royal North Shore Hospital, St Leonards, New South Wales, Australia.
Johnson
David W
DW
Vesey
David A
DA
Pollock
Carol A
CA
Chen
Xinming
X
D000469:Alkaline Phosphatase / Q000378:metabolism; D018929:Cell Culture Techniques; D002469:Cell Separation / Q000379:methods*; D048430:Cell Shape; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient / Q000379:methods; D004847:Epithelial Cells / Q000737:chemistry / Q000201:enzymology / Q000648:ultrastructure; D006801:Humans; D007633:Keratins / Q000032:analysis; D007687:Kidney Tubules, Proximal / Q000737:chemistry / Q000166:cytology* / Q000201:enzymology / Q000648:ultrastructure; D008871:Microvilli / Q000201:enzymology / Q000648:ultrastructure; D011205:Povidone; D012822:Silicon Dioxide; D017923:Sodium-Hydrogen Exchangers / Q000378:metabolism; D019108:Tight Junctions / Q000648:ultrastructure; D005723:gamma-Glutamyltransferase / Q000378:metabolism
D017923:Sodium-Hydrogen Exchangers; C016039:Percoll; D007633:Keratins; D012822:Silicon Dioxide; D005723:gamma-Glutamyltransferase; D000469:Alkaline Phosphatase; D011205:Povidone