GSM8565385: Unsorted and Lin- Sorted from ADM-adjacent mouse FBR capsule; Mus musculus; RNA-Seq
C57BL/6J mice underwent dorsal implantation with sterilized 0.2 mL PDMS implants.Mouse capsules were harvested at POD 28. Capsules were mechanically digested with sharp dissecting scissors (n = 6 per condition). Tissue was then added to an enzymatic digestion consisting of Collagenase II (ThermoFisher, Cat:17101015) and IV (ThermoFisher, Cat:17104019) in DMEM-F12 (GIBCOTM, Fischer Scientific, Hampton, NH). Samples were added to an orbital shaker at 150 rpm for 90 minutes at 37ºC. FACS buffer was added to quench the digest and samples were filtered through 70 µm cell strainers. Samples were then centrifuged at 1500 for 5 minutes at 4°C. For samples that were sorted, cells were counted and resuspended in FACS buffer. Primary antibodies were applied, and cells were stained in the dark with gentle agitation for 30 min. Cells were then washed thoroughly in FACS buffer. A BD II FACS Aria machine was used for FACS sorting analysis. For lineage negative (Lin-) FACS analysis, the following primary antibodies were used: Tie2, CD45, CD31, CD324, CD326, Ter119. Performed at the Stanford Genomics Facility (SGF) using the 10x Genomics platform. The relevant protocols were followed per manufacturer's guidelines.