Estimation of podocyte number: A comparison of methods
Estimation of podocyte number: A comparison of methods
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DOI：
10.1111/j.1523-1755.2004.00787.x
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发表时间：
2004-08-01
影响因子：
19.6
通讯作者：
Bilous, RW
中科院分区：
文献类型：
--
作者：
White, KE;Bilous, RW
背景足细胞是肾脏疾病进展机制研究的焦点，因此每个肾小球足细胞的数量已成为一个非常感兴趣的参数。当计数足细胞时，实际计数的颗粒是细胞核。大多数已发表的估计足细胞数量的研究使用了Weibel和Gomez（1962）的方法。这是对细胞核的形状和大小的假设，因此具有固有的偏差。在我们的研究中，我们使用了一种更新的体视学方法-分割器/分离器-它不对细胞核的形状或大小进行假设，因此没有偏倚。我们开始比较这两种方法，在1型糖尿病患者和正常对照组中，以确定消除偏倚从而提高准确性是否对总体结果有任何影响。Weibel-Gomez方法从肾小球的单个切片估计细胞数，而分离器/分离器需要连续切片肾小球。如果在电子显微镜照片上进行韦伯-戈麦斯估计，则两种方法获得的平均值之间无显著差异。然而，所有患者组的总体方差均较高，与所采用的方法无关。虽然分离器/分馏器是足细胞数量估计的理论金标准方法，但只要它们是从电子显微照片中获得的，则可以通过韦伯-戈麦斯方法获得可比的估计值。因此，可用的技术资源可决定所采用方法的选择。研究者应该意识到估计值的高度可变性，特别是当试图检测足细胞数量的微小变化时。
Background. The podocyte is the focus of much research into the mechanisms of renal disease progression, and the number of podocytes per glomerulus has thus become a parameter of much interest. When counting podocytes, the actual particle counted is the cell nucleus. The majority of published studies estimating podocyte number have used the method of Weibel and Gomez (1962). This makes assumptions about the shape and size of the cell nuclei and therefore has an inherent bias. In our studies we have used a more recent stereologic method-the disector/fractionator- that makes no assumptions about the shape or size of the cell nuclei and is therefore free of bias.Methods. We set out to compare the two methods, in both type 1 diabetic patients and normal controls, to determine whether eliminating bias and thus improving accuracy had any effect on the overall results. The Weibel-Gomez method estimates cell number from a single section through the glomerulus, whereas the disector/fractionator requires the glomerulus to be serially sectioned.Results. There was no significant difference between mean values obtained by the two methods, providing that the Weibel-Gomez estimate was performed on electron micrographs. However, the overall variance was high for all groups of patients, independent of the method employed.Conclusion. Although the disector/fractionator is the theoretic gold standard method for podocyte number estimation, comparable estimates can be obtained by the Weibel-Gomez method provided they are made from electron micrographs. Thus the technical resources available may determine the choice of method employed. Investigators should be aware of the high degree of variability in the estimate, particularly when trying to detect small changes in podocyte number.