Glomeruli Isolation: A Technique to Obtain Intact Glomeruli from Murine Kidney
내레이션 대본
In vertebrate kidneys, the nephrons consist of multiple glomeruli – a dense network of blood vessels – which play a primary role in the filtration of waste from the body.
To isolate glomeruli, begin by taking a freshly dissected mouse kidney. Use forceps to peel off the renal capsule from the kidney and place the clean kidney in a new culture dish. Now, position the dish over ice to avoid kidney degradation. Mince the kidney into small pieces.
Next, resuspend the minced kidney pieces in a suitable buffer solution. Filter the tissue suspension through a large-sized mesh filter and collect the flowthrough in a fresh dish. Repeat this step a few more times while progressively decreasing mesh size every time, till glomeruli dissociate from the kidney fragments.
Finally, wash the filter with the desired culture media to collect the smaller entrapped glomeruli while the kidney fragments remain in the filtrate. Transfer the glomeruli onto an ultra-low attachment plate. Visualize the plate using an inverted light microscope.
With the help of a micropipette, capture a single glomerulus and transfer it to a fresh well. Supplement the well with culture media and use the isolated glomerulus for further analysis.
