scRNA-Seq CD25+ intraperitoneal immune cells C57BL/6 mice implanted with engineered mAb expressing cells encapsulated in SLG20 alginate
Mouse intra-peritoneal washes and fat pads were procured two weeks after surgical implantation of Z2A19 or SLG20 capsules containing ADC26.2 cells, or sham surgeries. Five animals were collected for each condition, with final samples including mice with sham surgery, mice implanted with Z2A19, and mice implanted with SLG20. The peritoneal lavage, fat pads, and the explanted capsules were combined and converted into single cells by using the mouse fat protocol from the Miltenyi adipose tissue dissociation kit (catalog number: 130-105-808). Afterwards, cells were washed with phosphate-buffered saline (PBS) and were subsequently stained with Alexa Fluor 488 Anti-CD45 antibody [MRC OX-1] (Abcam catalog number: ab256254) for 1 hour. Cells were subsequently washed with PBS and resuspended in FACS buffer. Cells were then stained with NucBlue (Invitrogen catalog number: R37606) to assess for viability and run on an Aria III machine to collect roughly 500,000 viable cells per condition. A Chromium instrument by 10x Genomics X was used on 20,000 cells with 20,000 reads per cell, totaling 400M reads.
SUB16358338
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