Abstract:

Abstract Objective To describe a double fluorescent reporter mouse model to establish optimized protocol for GFP labeled podocyte isolation. Methodology: Based on fluorescent reporter transgenic mouse model together with modified extraction protocol, we demonstrated stable method to isolate podocyte using Fluorescence-activated cell sorting(FACS). Result:The efficient single cell dissociation to yield more than 300,000 purified podocytes per mouse were obtained allowing for global, unbiased downstream applications. Efficiency and purity could be monitored by evaluating sorted cells under a fluorescence microscope. Conclusion: RT-PCR analysis revealed that podocyte specific gene were highly enriched in GFP+ fraction, which was sufficient for further transcriptome profiling as well as quantitative proteomic analysis in omics approach.

Key words: FACS, reporter mice, podocyte