Abstract: FR-PO961
RNA Sequencing in Proximal Tubule Cells Reveals Lack of Expression of Proximal Tubule Markers
Session Information
- Pathology and Lab Medicine: Basic
November 08, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
Abstract Time: 10:00 AM - 12:00 PM
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 Symbol | Slc5a1 | Slc5a2 | Slc9a3 | Slc9a3r1 | Slc34a1 | Slc34a3 | LRP2 | Cldn1 | Ocln | Pth1r | Avpr2 |
| Common Name | SGLT-1 | SGLT-2 | NHE-3 | NHERF1 | NpT2a | NpT2c | Megalin | Claudin-1 | Occludin | PTH Receptor | ADH Receptor |
| HKC-8 | 23754 | 12420 | 11915 | 915 | 23101 | 21580 | 21088 | 7427 | 5404 | 10798 | 21540 |
| HKC-11 | 23754 | 14916 | 11222 | 1132 | 20077 | 23361 | 23028 | 7111 | 5166 | 10876 | 13060 |
| HPTC-05-LTR | 26753 | 11907 | 11703 | 997 | 26394 | 22534 | 25448 | 2696 | 9864 | 14722 | 25600 |
| HPTC-05-CLA | 27910 | 16403 | 11539 | 1124 | 22617 | 25157 | 25060 | 3526 | 11059 | 16557 | 25237 |
| HK-2 | 21749 | 13130 | 12915 | 2924 | 19159 | 21019 | 21016 | 1473 | 15373 | 13575 | 26872 |
| LLC-PK1 | 6036 | 7352 | 12612 | 455 | 3799 | 8342 | 2051 | 291 | 1371 | 14205 | 5507 |
| Rat-SHR | 16226 | 12143 | 15823 | 532 | 15830 | 14207 | 27648 | 2087 | 2424 | 13350 | 12977 |
| OK-WT | 683 | 6624 | 9106 | 743 | 1088 | 5677 | 2188 | 1209 | 12708 | 783 | No Expression |
| OK-H | 13464 | 3727 | 9886 | 2145 | 6831 | 13461 | 4829 | 1279 | 12135 | 2064 | No Expression |
Funding
- Other NIH Support