Abstract: FR-PO945
Spatial and Sex-Related Cell Diversity in the Adult Mouse Kidney Through scRNA-Seq Profiling
Session Information
- Development, Stem Cells, Regenerative Medicine - II
October 26, 2018 | Location: Exhibit Hall, San Diego Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Development, Stem Cells, and Regenerative Medicine
- 501 Development, Stem Cells, and Regenerative Medicine: Basic
Authors
- Ransick, Andrew, Broad-CIRM Center for Regenerative Med.& Stem Cell Research, Keck School of Med. of USC, Los Angeles, California, United States
- Kim, Albert D., University of Southern California, Los Angeles, California, United States
- McMahon, Andrew P., Keck School of Medicine of the University of Southern California, Los Angeles, California, United States
Background
Applying single cell RNA Sequencing (scRNA-seq) methodology to adult mouse kidneys facilitates examination of the gene expression profiles of individual kidney cell-types in high resolution. Recent studies have shown the power of this approach for adult mouse whole kidney (Han et al., Cell; Park et al, Science). By incorporating zonal dissection of tissue samples and both sexes, we are generating a high resolution map of kidney cell types that integrates spatial organization and sex-related differences.
Methods
We profiled adult kidneys from two male and female wildtype C57B6/J strain mice. Prior to tissue dissociation, kidneys were dissected along the corticomedullary axis (CMA) to isolate pieces containing exclusively cortex, outer medulla or inner medulla/papilla. Tissue dissociation at 12 C utilized cold active proteases to reduce stress responses. scRNA-seq profiling used 10XGenomics Chromium platform. PCA and identification of variably expressed genes executed with Seurat v2.3 package in RStudio.
Results
We obtained 38,039 transcriptional profiles from four mice representing all expected cell types of the adult kidney, though podocytes were underrepresented. Initial analysis has focused on identifying lineage, zonal and sex differences. We observe expected zonal clustering for known cell types. Although within specific lineages distributed along the CMA, we find regional diversity in profiles and can identify cell types excluded from published scRNA-seq studies. A marked sex diversity was observed in specific cell populations, confirming and extending known sex-related differences in kidney gene expression. Our zonal analysis also facilitates mapping regional diversity amongst the associated cell populations (eg. vascular, mesenchyme, immune-related).
Conclusion
These data generate a solid foundation for building a high resolution map of cell diversity in the adult male and female mouse kidney. We will discuss our progress towards this goal and review new insights that have come from scRNA-seq analysis of specific kidney lineages.
Funding
- NIDDK Support