Table 1 Protocol for direct dissociation of a whole kidney into a cell suspension

1.

Prepare sterile dissociation buffer (4°C): PBS 1×, 0.5% bovine serum albumin, and 2 mM EDTA.

2.

Immerse the kidney in 1 mL of this dissociation buffer

3.

Remove the capsule and roughly chop the kidney with a surgical blade

4.

Transfer the solution obtained into GentleMACS C-tubes.

5.

Use the brain_03 and then the spleen_04 program of the GentleMACS dissociator.

6.

Pass the solution obtained through a 30-μm sieve.

7.

Rinse with 4 mL of dissociation buffer.

8.

Centrifuge at 500 g for10 minutes.

9.

Discard the supernatant.

10

Resuspend cell pellet in 180 μL of dissociating buffer.