Conditionally immortalized human podocyte cell lines established from urine.
Evidence suggests that loss of podocytes into urine contributes to development of glomerular diseases; shed podocytes are frequently viable and proliferate in culture conditions. To determine the phenotypic characteristics of viable urinary cells derived from human subjects, we established long-term urinary cell culture from two patients with focal segmental glomerulosclerosis and two healthy volunteers, via transformation with the thermosensitive SV40 large T antigen (U19tsA58) together with human telomerase (hTERT). Characterization of arbitrarily selected two clonal cell lines from each human subject was carried out. mRNA expression for the podocyte markers synaptopodin, nestin, and CD2AP were detected in all eight clones. Podocin mRNA was absent from all eight clones. The expression of nephrin, Wilms' tumor 1 (WT1), and podocalyxin mRNA varied among the clones, which may be due to transformation and/or cloning. These results suggest that podocyte cell lines can be established consistently from human urine. The generation of podocyte cell lines from urine of patients and healthy volunteers is novel and will help to advance studies of podocyte cell biology. Further improvements in the approaches to cell transformation and/or cell culture techniques are needed to allow cultured podocytes to fully reproduce in vivo characteristics.
Sakairi
Toru
T
Kidney Disease Section, Kidney Disease Branch, National Institute of Diabetes, Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892-1268, USA.
Abe
Yoshifusa
Y
Kajiyama
Hiroshi
H
Bartlett
Linda D
LD
Howard
Lilian V
LV
Jat
Parmijit S
PS
Kopp
Jeffrey B
JB
D000328:Adult; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D015415:Biomarkers / Q000378:metabolism; D016022:Case-Control Studies; D002454:Cell Differentiation; D002461:Cell Line, Transformed; D049109:Cell Proliferation; D002469:Cell Separation; D002470:Cell Survival; D002472:Cell Transformation, Viral / Q000235:genetics*; D002999:Clone Cells; D005260:Female; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D005819:Genetic Markers; D005923:Glomerulosclerosis, Focal Segmental / Q000235:genetics / Q000473:pathology* / Q000652:urine; D006801:Humans; D008297:Male; D008875:Middle Aged; D010641:Phenotype; D050199:Podocytes / Q000378:metabolism / Q000473:pathology*; D012333:RNA, Messenger / Q000378:metabolism; D019098:Telomerase / Q000235:genetics*; D013997:Time Factors; D014556:Urine / Q000166:cytology
D000952:Antigens, Polyomavirus Transforming; D015415:Biomarkers; D005819:Genetic Markers; D012333:RNA, Messenger; C509186:TERT protein, human; D019098:Telomerase