Python automated tissue dissociation system

We dissected out the testes of the 40 days group, rinsed them using cold phosphate‐buffered saline (PBS; HyClone, Logan, UT, USA), and transferred them immediately into ice‐cold GEXSCOPE™ Tissue Preservation Solution (Singleron Biotechnologies, Cheshire, CT, USA). The testes were then rinsed thrice using Hanks Balanced Salt Solution (HBSS), digested in 2 mL of GEXSCOPE™ Tissue Dissociation Solution (Singleron Biotechnologies) in a Singleron PythoN™ Automated Tissue Dissociation System for 15 min at 28°C, subjected to centrifugation for 5 min at 500 × g, and suspended in PBS. Trypan blue (Sigma‐Aldrich, St. Louis, MO, USA) was used to stain the samples, whose viability was determined under a microscope.

The intestine of the control and CLP-treated group from WT and Sting1^−/− mice were removed at the indicated time (n = 3 for each group). The fresh tissues were washed with Hank’s balanced salt solution (HBSS) three times and then minced into 1–2 mm pieces. The tissue pieces were digested with 2 mL CelLiveTM Tissue Dissociation Solution (Singleron) by Singleron PythoN™ Automated Tissue Dissociation System (Singleron) at 37 °C for 15 min. Afterward, cells were incubated with red blood cell lysis buffer for another 10 min. The solution was then centrifuged at 500 g for 5 min and suspended softly with PBS. Finally, the samples were stained with trypan blue (Sigma, United States), and the cellular viability was evaluated microscopically.