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Abstract: FR-PO961

RNA Sequencing in Proximal Tubule Cells Reveals Lack of Expression of Proximal Tubule Markers

Session Information

Category: Pathology and Lab Medicine

  • 1601 Pathology and Lab Medicine: Basic

Authors

  • Khundmiri, Syed J., Howard University College of Medicine, Washington, District of Columbia, United States
  • Chen, Lihe, NIH, Bethesda, Maryland, United States
  • Lederer, Eleanor D., University of Louisville; Robley Rex VA Medical Center, Louisville, Kentucky, United States
  • Knepper, Mark A., National Heart Lung and Blood Institute, Kensington, Maryland, United States
Background

There are close to 12 different immortalized cell lines available commercially or from other sources that are being used as models of renal proximal tubule cells. There is strong need to characterize these cells so as to establish their lineage and help the scientific community at large to make decisions regarding their suitability in experiments. Our goal is to characterize the cell lines using RNA-Seq.

Methods

To accomplish this goal, we grew 5 different human cell lines, one mouse, three rat, and 2 opossum kidney cell lines on Transwell filters using the recommended culture media. RNA-Seq was performed using Illumina kits following the manufacturer's protocol. The RNA-Seq was compared to a mouse S1 proximal tubule data. The data are presented as expression rank with the rank above 8000 as negative expression. To validate the data a native S1 proximal tubule cell line was used as a positive control.

Results

Results are shown in the Table.

Conclusion

Our data suggest that the available human and rat kidney tubule cells are epithelial cells (express occludin and claudins) but not of proximal tubule origin (lack sodium-dependent transporters, megalin and PTH receptor). Although, LLC-PK1 cells express the proximal tubule specific proteins, they also express markers of cortical tubule cells. These are therefore a mixed population and not specific proximal tubule cells. The opossum kidney cells express all the markers of proximal tubule cells and thus are the only cell of proximal tubule origin.

Expression of proximal tubule markers
Gene SymbolSlc5a1Slc5a2Slc9a3Slc9a3r1Slc34a1Slc34a3LRP2Cldn1OclnPth1rAvpr2
Common Name
SGLT-1SGLT-2NHE-3NHERF1NpT2aNpT2cMegalinClaudin-1OccludinPTH ReceptorADH Receptor
HKC-8
237541242011915915231012158021088742754041079821540
HKC-112375414916112221132200772336123028711151661087613060
HPTC-05-LTR267531190711703997263942253425448269698641472225600
HPTC-05-CLA27910164031153911242261725157250603526110591655725237
HK-221749131301291529241915921019210161473153731357526872
LLC-PK160367352126124553799834220512911371142055507
Rat-SHR162261214315823532158301420727648208724241335012977
OK-WT68366249106743108856772188120912708783No Expression
OK-H1346437279886214568311346148291279121352064No Expression

Funding

  • Other NIH Support