GFR measured using DBS in animal models – Laboratorio de Función Renal

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Measurement of renal function in experimental animal models using the DBS (Dried Blood Spot) technique

As in humans, in our laboratory we have developed a simple and reliable method for the measurement of renal function by plasma clearance of iohexol in experimental animal models using the technique of dried blood spot (DBS).

In mice, the method consists of a single intravenous injection through the lateral vein of the tail of the animal with a solution of iohexol in physiological serum (1:10) (Figure 1). The animal should be lightly sedated to avoid pain and/or discomfort and for the injection to be accurate.

Figure 1.A Animal model: mouse; Figure 1.B Intravenous injection into the mouse tail

Then, with the animal conscious and without movement restriction, blood samples are taken from the tip of the tail of the animal with heparinized capillaries of fixed volume (5uL) and deposited on a special filter paper (Whatman 903, GE Healthcare) (Figure 2). The filter paper is left to dry 24 and then the iohexol is extracted and measured by liquid chromatography (HPLC) in the same way as in the human method.

Figure 2.A Take a 5ul heparinized capillary; Figure 2.B Blood extraction from the animal’s tail vein; Figure 2.C Placing the sample on special filter paper (Whatman 903, GE Healthcare); Figure 2.D Capillary completely empty

With this method, animal welfare is improved since a minimum volume of blood is required for the entire procedure. Controlling blood volume in small experimental animal procedures is crucial for reproducible and reliable results.
This method has been validated in mice^1,2, pigs³, sheeps⁴ and rats⁵.

Here you can find an explanatory video of the DBS method:

Scientific reference for the DBS method in animal models:

Rodriguez-Rodriguez AE, Luis-Lima S, Donate-Correa J, Diaz-Martin L, Arnau MR, Jimenez-Sosa A, et al. Iohexol plasma clearance simplified by Dried Blood Spot (DBS) sampling to measure renal function in conscious mice. Sci Rep. 2021;11(1):4591.
Luis-Lima S, Rodriguez-Rodriguez AE, Martin-Higueras C, Sierra-Ramos C, Carrara F, Arnau MR, et al. Iohexol plasma clearance, a simple and reliable method to measure renal function in conscious mice. Pflugers Arch. 2016;468(9):1587-94.
Rodriguez RR, Gonzalez-Bulnes A, Garcia-Contreras C, Elena Rodriguez-Rodriguez A, Astiz S, Vazquez-Gomez M, et al. The Iberian pig fed with high-fat diet: a model of renal disease in obesity and metabolic syndrome. Int J Obes (Lond). 2020;44(2):457-65
Luis-Lima S, Garcia-Contreras C, Vazquez-Gomez M, Astiz S, Carrara F, Gaspari F, et al. A Simple Method to Measure Renal Function in Swine by the Plasma Clearance of Iohexol. Int J Mol Sci. 2018;19(1).
Carrara F, Azzollini N, Nattino G, Corna D, Villa S, Cerullo D, et al. Simplified Method to Measure Glomerular Filtration Rate by Iohexol Plasma Clearance in Conscious Rats. Nephron. 2016;133(1):62-70.