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| Status |
Public on Apr 04, 2023 |
| Title |
Arsenite Exposure to Human Renal Progenitor Cells (HRTPT) Produces a Reversible Epithelial Mesenchymal Transition (EMT) |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Background The human kidney is known to possess renal progenitor cells (RPCs) that can assist in the repair of acute tubular injury. The RPCs are sparsely located as single cells throughout the kidney. We recently generated an immortalized human renal progenitor cell line (HRTPT) that co-expresses PROM1/CD24 and expresses features expected on a RPCs. This included the ability to form nephrospheres, differentiate on the surface of Matrigel, and to undergo adipogenic, neurogenic, and osteogenic differentiation. These cells were used in the present study to determine how the cells would respond when exposed to a nephrotoxin. Arsenite (iAS) was chosen as the nephrotoxin since the kidney is susceptible to this toxin and there is evidence for its involvement in renal disease. Methods gene expression profiles when the cells were exposed to iAS for 3, 8, 10 passages (subcultured at 1:3 ratio) identified a shift in from the control unexposed cells. The cells exposed to iAS for 8 passages were then referred with growth media containing no iAS and within 2 passages the cells returned to an epithelial morphology with strong agreement in differential gene expression between control and cells recovered from iAS exposure. Results Results shows within 3 serial passages of the cells exposed to iAS there was a shift in morphology from an epithelial to a mesenchymal phenotype. EMT was suggested based on an increase in known mesenchymal markers. Conclusion We found RPCs can undergo EMT when exposed to a nephrotoxin and undergo MET when the agent is removed from the growth media.
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| Overall design |
The isolation and serum-free culture conditions for the HRTPT cells has been previously described.10, 11 Confluent cultures of HRTPT cells were exposed to 4.5 µM iAS for 24 hrs and then subcultured at a 1:3 ratio in the continued presence of iAS until confluent. Following confluence, the cells were serially sub cultured again in the present of iAS until confluent. This was repeated for 10 serial passages. Additional cultures of iAS-exposed cells at passage 8 were subcultured into iAS-free growth media and continued in iAS-free media for 11 additional passages. Genome Explorations has completed a gene expression analysis of 18 samples using the Clariom™ D microarray to determine the effects of Arsenite treatment on renal proximal tubule cell gene expression.
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| Contributor(s) |
Singhal S, Shrestha S, Singhal S, Sens D |
| Citation(s) |
36982180 |
| |
| Submission date |
Oct 17, 2022 |
| Last update date |
Apr 05, 2023 |
| Contact name |
Sandeep K Singhal |
| E-mail(s) |
Sandeep.singhal@und.edu
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| Organization name |
University of North Dakota
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| Department |
Pathology
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| Street address |
1301 N Columbia Rd Stop 9037
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| City |
Grand Forks |
| State/province |
NORTH DAKOTA |
| ZIP/Postal code |
58202 |
| Country |
USA |
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| Platforms (1) |
| GPL23126 |
[Clariom_D_Human] Affymetrix Human Clariom D Assay [transcript (gene) version] |
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| Samples (18)
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| Relations |
| BioProject |
PRJNA891267 |
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