Abstract: FR-PO181
Primary Cultures from Injured and Healthy Mouse Kidneys: A Comparative Transcriptomic Analysis
Session Information
- AKI: Mechanisms
October 25, 2024 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Acute Kidney Injury
- 103 AKI: Mechanisms
Authors
- López-Cayuqueo, Karen I., Medizinische Hochschule Hannover, Hannover, Niedersachsen, Germany
- Tsamo Tetou, Arnold, Medizinische Hochschule Hannover, Hannover, Niedersachsen, Germany
- Günzel, Dorothee, Charite Universitatsmedizin Berlin, Berlin, Berlin, Germany
- Schmidt-Ott, Kai M., Medizinische Hochschule Hannover, Hannover, Niedersachsen, Germany
Background
Primary cell cultures from mouse kidneys are widely used to investigate kidney physiology and pathophysiology. It is known culture systems are far from being an ideal model but there is still a need for in vitro systems to validate different hypotheses. Here, we prepared primary cell cultures of renal cortex from both healthy and injured kidneys, and we compare their transcriptomics landscape intending to determine whether cultured kidney cells from an injured kidney are representative of in vivo injury states.
Methods
Unilateral Ischemia-Reperfusion Injury was performed in C57BL/6N mice. After 21 days of reperfusion primary cell cultures were prepared from the renal cortex of both contralateral and injured kidneys. The cells were kept in culture for 6 days before being analyzed using bulk RNAseq and single-cell sequencing. Additionally, cell cultures were compared to uncultured kidney cells from contralateral and injured kidneys (from the same mouse).
Results
Cell cultures only partially resemble in vivo conditions. During culturing, the differences between cells from injured kidneys and those from healthy kidneys become less pronounced. Surprisingly, cultured renal cells from healthy kidneys adopt an injured kidney phenotype, detonated by the upregulation of kidney injury markers such as Havcr1 and Vcam1. Despite the similarities between healthy and injured cultured cells, it is possible to identify key differences at the single-cell level, such as the identification of additional clusters represented by monocyte and macrophage markers.
Conclusion
We demonstrated that primary cell cultures from healthy mouse renal cortex could be a suitable model to investigate injury-associated pathways since the culturing step induces in the cells a kidney injury-associated phenotype. However, culturing cells from an injured kidney potentially offers additional information, such as the presence of immune cells.
Funding
- Government Support – Non-U.S.