The TH1 cells are derived from primary human renal proximal tubule epithelial cells (RPTECs) immortalized by two lentiviral vectors carrying the human telomerase (hTERT) and the SV40 T antigen (Tag).
These cells can sustain long-term proliferation in culture, exhibit contact-inhibited, anchorage- and growth factor-dependent growth and do not form tumors in nude mice. TH1 cells express alkaline phosphatase and gamma-glutamyltranspeptidase, two brush-border enzymes expressed in RPTECs. TH1 cells also perform critical metabolic activities characteristic of renal proximal tubule cells, including active glucose transport and ammonia production and excretion.
Both the hTERT and Tag genes in the lentiviral vectors are flanked by loxP sites. Transient Cre expression in TH1 cells leads to efficient proviral deletion and upregulation of some renal specific activities. Removal of the genes for cell immortalization also significantly decreases the rate of cell proliferation and ultimately leads to cell senescence, suggesting that the cell cycle control remains intact in TH1 cells despite continuous passage of these cells in culture.
From the laboratory of Jiing-Kuan Yee, PhD, City of Hope.
Part of The Investigator's Annexe program.
|Product Type:||Cell Line|
|Cell Type:||Proximal tubule epithelial cells|
|Growth Conditions:||DMEM-HIGH Glucose (HyClone # SH30022.01) and 10% FBS (HyClone # SV30014.03)|
|Subculturing:||1 to 5 dilution every 5 days|
|Cryopreservation:||DMEM + 20% FBS + 10% DMSO|
TH1 Cell morphology.
(a) Primary RPTECs at passage 7. (b) Primary RPTECs at passage 7 stained for senescence-specific b-Gal expression. (c) TH1 cells at passage 30. (d) Immunofluorescent staining of cells in (c) for cytokeratin. (e) Cell proliferation assay. BrdU incorporation was measured in proliferating and confluent TH1 cells. The results were normalized to the cell number.
Adapted from: Kowolik CM, Liang S, Yu Y & Yee JK. Cre-mediated reversible immortalization of human renal proximal tubular epithelial cells. Oncogene, 23:5950-5957, (2004).
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