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1.  Using Stereologic Techniques for Podocyte Counting in the Mouse: Shifting the Paradigm 
American Journal of Nephrology  2011;33(Suppl 1):1-7.
Background
The podocyte serves the important function of maintaining the glomerular filtration barrier, and many studies report a decrease in podocyte number relative to the development of proteinuric states. However, there is significant inconsistency in the number of podocytes counted, possibly due to different counting methods. We previously counted the three glomerular cell types in the mouse kidney and showed that the fractionator/disector method is a close approximation of the exhaustive count or the gold standard method. In this study, we compared the commonly used model-based approach with the design-based approach to count podocytes in the db/m and db/db mouse and illustrate that the design-based approach, which uses the fractionator/disector method, provides an accurate determination of podocyte number.
Methods
In the design-based approach, toluidine blue-stained 1-μm-thick sections from glutaraldehyde perfusion-fixed kidneys were used (n = 15) with the fractionator/disector method. In the model-based approach, WT-1-immunolabeled podocyte nuclei in 3- to 4-μm-thick formalin-fixed paraffin-embedded sections of the same kidneys were counted with the Weibel-Gomez method. Glomerular volume was determined for each method.
Results
We discovered that the fractionator/disector method counted 89 ± 10 podocytes compared to the Weibel-Gomez method, which counted 137 ± 38 podocytes and overestimated podocyte number by 54% (p < 0.05). In addition, glomerular volume (231 ± 52 × 103 vs. 192 ± 64 × 103 μm3) was significantly underestimated by 17% (p < 0.0002). Moreover, the model-based approach was more time consuming.
Conclusion
We conclude that the fractionator/disector method offers an unbiased and efficient determination of podocyte counts.
doi:10.1159/000327564
PMCID: PMC3121548  PMID: 21659728
db/db mouse; Design-based approach; Fractionator/disector method; Glomerular volume; Podocyte number; Stereology; Weibel-Gomez method
2.  Podocyte Number in the Maturing Rat Kidney 
American Journal of Nephrology  2010;33(1):91-96.
Background/Aims
The podocyte is an important cell for maintaining the normal structure and function of the glomerulus. In recent years much attention has been given to the number of podocytes in glomeruli. During this time there has been a debate as to whether podocytes can divide after the capillary-loop stage of development. The aim of this study was to use an unbiased counting method to determine if podocyte number increases after the capillary-loop stage of development.
Methods
The fractionator/disector method was used to count podocytes in glomeruli from rats aged 1 day, 5, 12, and 24 weeks. Glomerular volume was also measured with the unbiased Cavalieri principle and used to calculate the density of podocytes per glomerulus.
Results
The number of podocytes did not increase from the capillary-loop stage of glomerular development to 24 weeks of age. Glomerular volume increased 3.6-fold during this time, which resulted in a decrease of podocyte density as the rats aged.
Conclusion
The study documents that the number of podocytes is stable after the capillary-loop stage of glomerular development. The data does not confirm but adds evidence that podocytes do not divide from the capillary-loop stage of glomerular development to 24 weeks of age in the normal rat.
doi:10.1159/000322701
PMCID: PMC3030545  PMID: 21196721
Fractionator/disector; Glomerular volume; Podocyte number; Rat; Stereology

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