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An Efficient Sieving Method to Isolate Intact Glomeruli from Adult Rat Kidney

An Efficient Sieving Method to Isolate Intact Glomeruli from Adult Rat Kidney

Published on: November 1, 2018

Kinetics of Label Retaining Cells in the Developing Rat Kidneys.

Jianwen Wang^1,2, Guiting Lin², Amjad Alwaal²

¹Department of Urology, Beijing ChaoYang Hospital, Capital Medical University, 8 Gongtinanlu, Beijing, 100020, China.

|December 10, 2015

Summary

This study introduces a new method using 5-ethynyl-2-deoxyuridine (EdU) to track kidney stem cells. EdU-labeled label-retaining cells (LRCs) were found in glomeruli and tubules, showing potential for kidney regeneration.

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Area of Science:

Nephrology
Stem Cell Biology
Developmental Biology

Background:

Kidneys possess limited regenerative capacity and diverse cell lineages.
Traditional BrdU labeling for kidney stem cells is hindered by antibody accessibility issues.
A novel 5-ethynyl-2-deoxyuridine (EdU) labeling method was developed to overcome these limitations.

Purpose of the Study:

To introduce and validate a new cell labeling method using EdU for kidney stem cell identification.
To investigate the dynamic changes and localization of EdU-labeled cells over time in developing kidneys.
To assess the co-expression of potential stem/progenitor markers with EdU-labeled cells.

Main Methods:

Newborn rats were injected with EdU.
Kidneys were harvested at multiple time points (1 day to 6 weeks).
Immunofluorescence staining was used to detect EdU and cellular markers.

Main Results:

EdU-labeled label-retaining cells (LRCs) decreased from day 1 to week 6.
LRC distribution differed between renal cortex and papilla over time.
At 6 weeks, LRCs were found in glomeruli (expressing podocyte/endothelial markers) and tubules (expressing Nestin/vascular markers).

Conclusions:

The EdU labeling method effectively tracks kidney stem/progenitor cells.
Long-term LRCs in glomeruli express undifferentiated markers, suggesting a role in podocyte and endothelial maintenance.
Further research is needed to fully characterize EdU-LRCs and their regenerative potential in the kidney.